Dual-acting benzoimidazole antihypertensive agents

ABSTRACT

The invention is directed to compounds having the formula: 
     
       
         
         
             
             
         
       
         
         
           
             wherein: Ar, r, n, X, R 2-3  and R 5-7  are as defined in the specification, and pharmaceutically acceptable salts thereof. These compounds have AT 1  receptor antagonist activity and neprilysin inhibition activity. The invention is also directed to pharmaceutical compositions comprising such compounds; methods of using such compounds; and process and intermediates for preparing such compounds.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No. 60/933,207, filed on Jun. 5, 2007; the entire disclosure of which is incorporated herein by reference in its entirety.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to novel compounds having angiotensin II type 1 (AT₁) receptor antagonist activity and neprilysin-inhibition activity. The invention also relates to pharmaceutical compositions comprising such compounds, processes and intermediates for preparing such compounds and methods of using such compounds to treat diseases such as hypertension.

2. State of the Art

The aim of antihypertensive therapy is to lower blood pressure and prevent hypertension-related complications such as myocardial infarction, stroke, and renal disease. For patients with uncomplicated hypertension (that is, no risk factors, target organ damage, or cardiovascular disease), it is hoped that reducing blood pressure will prevent development of cardiovascular and renal comorbidities, conditions that exist at the same time as the primary condition in the same patient. For those patients with existing risk factors or comorbidities, the therapeutic target is the slowing of comorbid disease progression and reduced mortality.

Physicians generally prescribe pharmacological therapies for patients whose blood pressure cannot be adequately controlled by dietary and/or lifestyle modifications. Commonly used therapeutic classes act to promote diuresis, adrenergic inhibition, or vasodilation. A combination of drugs is often prescribed, depending upon what comorbidities are present.

There are five common drug classes used to treat hypertension: diuretics, which include thiazide and thiazide-like diuretics such as hydrochlorothiazide, loop diuretics such as furosemide, and potassium-sparing diuretics such as triamterene; β₁ adrenergic receptor blockers such as metoprolol succinate and carvedilol; calcium channel blockers such as amlodipine; angiotensin-converting enzyme (ACE) inhibitors such as captopril, benazepril, enalapril, enalaprilat, lisinopril, quinapril, and ramipril; and AT₁ receptor antagonists, also known as angiotensin II type 1 receptor blockers (ARBs), such as candesartan cilexetil, eprosartan, irbesartan, losartan, olmesartan medoxomil, telmisartan, and valsartan. Combinations of these drugs are also administered, for example, a calcium channel blocker (amlodipine) and an ACE inhibitor (benazepril), or a diuretic (hydrochlorothiazide) and an ACE inhibitor (enalapril). All of these drugs, when used appropriately, are effective in the treatment of hypertension. Nevertheless, both efficacy and tolerability should be further improved in new drugs targeting hypertension. Despite the availability of many treatment options, the recent National Health And Nutrition Examination Survey (NHANES) demonstrated that only about 50% of all treated patients with hypertension achieve adequate blood pressure control. Furthermore, poor patient compliance due to tolerability issues with available treatments further reduces treatment success.

In addition, each of the major classes of antihypertensive agents have some drawbacks. Diuretics can adversely affect lipid and glucose metabolism, and are associated with other side effects, including orthostatic hypotension, hypokalemia, and hyperuricemia. Beta blockers can cause fatigue, insomnia, and impotence; and some beta blockers can also cause reduced cardiac output and bradycardia, which may be undesirable in some patient groups. Calcium channel blockers are widely used but it is debatable as to how effectively these drugs reduce fatal and nonfatal cardiac events relative to other drug classes. ACE inhibitors can cause coughing, and rarer side effects include rash, angioedema, hyperkalemia, and functional renal failure. AT₁ receptor antagonists are equally effective as ACE inhibitors but without the high prevalence of cough.

Neprilysin (neutral endopeptidase, EC 3.4.24.11) (NEP), is an endothelial membrane bound Zn²⁺ metallopeptidase found in many tissues, including the brain, kidney, lungs, gastrointestinal tract, heart, and peripheral vasculature. NEP is responsible for the degradation and inactivation of a number of vasoactive peptides, such as circulating bradykinin and angiotensin peptides, as well as the natriuretic peptides, the latter of which have several effects including vasodilation and diuresis. Thus, NEP plays an important role in blood pressure homeostasis. NEP inhibitors have been studied as potential therapeutics, and include thiorphan, candoxatril, and candoxatrilat. In addition, compounds have also been designed that inhibit both NEP and ACE, and include omapatrilat, gempatrilat, and sampatrilat. Referred to as vasopeptidase inhibitors, this class of compounds are described in Robl et al. (1999) Exp. Opin. Ther. Patents 9(12): 1665-1677.

There may be an opportunity to increase anti-hypertensive efficacy when combining AT₁ receptor antagonism and NEP inhibition, as evidenced by AT₁ receptor antagonist/NEP inhibitor combinations described in WO 9213564 to Darrow et al (Schering Corporation); US20030144215 to Ksander et al.; Pu et al., Abstract presented at the Canadian Cardiovascular Congress (October 2004); and Gardiner et al. (2006) JPET 319:340-348; and WO 2007/045663 (Novartis AG) to Glasspool et al.

Recently, WO 2007/056546 (Novartis AG) to Feng et al. has described complexes of an AT₁ receptor antagonist and a NEP inhibitor, where an AT₁ receptor antagonist compound is non-covalently bound to a NEP inhibitor compound, or where the antagonist compound is linked via non-covalent bonding to the inhibitor compound.

In spite of the advances in the art, there remains a need for a highly efficacious monotherapy with multiple mechanisms of action leading to levels of blood pressure control that can currently only be achieved with combination therapy. Thus, although various hypertensive agents are known, and administered in various combinations, it would be highly desirable to provide compounds having both AT₁ receptor antagonist activity and NEP inhibition activity in the same molecule. Compounds possessing both of these activities are expected to be particularly useful as therapeutic agents since they would exhibit antihypertensive activity through two independent modes of action while having single molecule pharmacokinetics.

In addition, such dual-acting compounds are also expected to have utility to treat a variety of other diseases that can be treated by antagonizing the AT₁ receptor and/or inhibiting the NEP enzyme.

SUMMARY OF THE INVENTION

The present invention provides novel compounds that have been found to possess AT₁ receptor antagonist activity and neprilysin (NEP) enzyme inhibition activity. Accordingly, compounds of the invention are expected to be useful and advantageous as therapeutic agents for treating conditions such as hypertension and heart failure.

One aspect of the invention is directed to a compound of formula I:

wherein:

r is 0, 1 or 2;

Ar is an aryl group selected from:

R¹ is selected from —COOR″, —NHSO₂R^(1b), —SO₂NHR^(1d), —SO₂OH, —C(O)NH—SO₂R^(1c), —P(O)(OH)₂, —CN, —OCH(R^(1c))—COOH, tetrazol-5-yl,

R^(1a) is H, —C₁₋₆alkyl, —C₁₋₃alkylenearyl, —C₁₋₃alkyleneheteroaryl, —C₃₋₇cycloalkyl, —CH(C₁₋₆alkyl)OC(O)R^(1aa), —C₀₋₆alkylenemorpholine,

R^(1aa) is —O—C₁₋₆alkyl, —O—C₃₋₇cycloalkyl, —NR^(1ab)R^(1ac), or —CH(NH₂)CH₂COOCH₃; R^(1ab) and R^(1ac) are independently selected from H, —C₁₋₆alkyl, and benzyl, or are taken together as —(CH₂)₃₋₆—; R^(1b) is R^(1c) or —NHC(O)R^(1c); R^(1c) is —C₁₋₆alkyl, —C₀₋₆alkylene-O—R^(1ca), —C₁₋₅alkylene-NR^(1cb)R^(1cc), —C₀₋₆alkylenearyl or —C₀₋₆alkyleneheteroaryl; R^(1ca) is H, —C₁₋₆alkyl, or —C₁₋₆alkylene-O—C₁₋₆alkyl; R^(1cb) and R^(1cc) are independently selected from H and —C₁₋₆alkyl, or are taken together as —(CH₂)₂—O—(CH₂)₂— or —(CH₂)₂—N[C(O)CH₃]-(CH₂)₂—; R^(1d) is H, R^(1c), —C(O)R^(1c), or —C(O)NHR^(1c); R^(1e) is —C₁₋₄alkyl or aryl;

n is 0, 1, 2 or 3;

each R² is independently selected from halo, —NO₂, —C₁₋₆alkyl, —C₂₋₆alkenyl, —C₃₋₆cycloalkyl, —CN, —C(O)R^(2a), —C₀₋₅alkylene-OR^(2b), —C₀₋₅alkylene-NR^(2c)R^(2d), —C₀₋₃alkylenearyl, and —C₀₋₃alkyleneheteroaryl; where R^(2a) is selected from H, —C₁₋₆alkyl, —C₃₋₆cycloalkyl, —OR^(2b), and —NR^(2c)R^(2d); R^(2b) is selected from H, —C₁₋₆alkyl, —C₃₋₆cycloalkyl, and —C₀₋₁alkylenearyl; and R^(2a) and R^(2d) are independently selected from H, —C₁₋₄alkyl, and —C₀₋₁alkylenearyl;

R³ is selected from —C₁₋₁₀alkyl, —C₂₋₁₀alkenyl, —C₃₋₁₀alkynyl, —C₀₋₃alkylene-C₃₋₇cycloalkyl, —C₂₋₃alkenylene-C₃₋₇cycloalkyl, —C₂₋₃alkynylene-C₃₋₇cycloalkyl, —C₀₋₅alkylene-NR^(3a)—C₀₋₅alkylene-R^(3b), —C₀₋₅alkylene-O—C₀₋₅alkylene-R^(3b), —C₀₋₅alkylene-S—C₁₋₅alkylene-R^(3b), and —C₀₋₃alkylenearyl; where R^(1a) is selected from H, —C₁₋₆alkyl, —C₃₋₇cycloalkyl, and —C₀₋₃alkylenearyl; and R^(3b) is selected from H, —C₁₋₆alkyl, —C₃₋₇cycloalkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl, and aryl;

X is —C₁₋₁₂alkylene-, where at least one —CH₂— moiety in the alkylene is replaced with a —NR^(4a)—C(O)— or —C(O)—NR^(4a)— moiety, where R^(4a) is selected from H, —OH, and —C₁₋₄alkyl;

R⁵ is selected from —C₀₋₃alkylene-SR^(5a), —C₀₋₃alkylene—C(O)NR^(5b)R^(5c), —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d), —NH—C₀₋₁alkylene-P(O)(OR^(5e))₂, —C₀₋₃alkylene-P(O)OR^(5e)R^(5f), C₀₋₂alkylene-CHR^(5g)—COOH, —C₀₋₃alkylene—C(O)NR^(5h)—CHR^(5i)—COOH, and —C₀₋₃alkylene-S—SR^(5j); where R^(5a) is H or —C(O)—R^(5aa); R^(5aa) is —C₁₋₆alkyl, —C₀₋₆alkylene-C₃₋₇cycloalkyl, -aminoC₄₋₇cycloalkyl, —C₀₋₆alkylenearyl, —C₀₋₆alkyleneheteroaryl, —C₀₋₆alkylenemorpholine, —C₀₋₆alkylenepiperazine—CH₃, —C₀₋₆alkylenepiperidine, —C₀₋₆alkylenepiperidine—CH₃, —CH[N(R^(5ab))₂]-aa where aa is an amino acid side chain, —C₀₋₆alkylene-CH[N(R^(5ab))₂]—R^(5ac), -2-pyrrolidine, -2-tetrahydrofuran, —C₀₋₆alkylene-OR^(5ab), —O—C₀₋₆alkylenearyl, —C₁₋₂alkylene-OC(O)—C₁₋₆alkyl, —C₁₋₂alkylene-OC(O)—C₀₋₆alkylenearyl, —O—C₁₋₂alkylene-OC(O)O—C₁₋₆alkyl, —C₁₋₄alkylene-COOH, or -arylene-COOH; R^(5ab) is independently H or —C₁₋₆alkyl; R^(5ac) is H, —C₁₋₆alkyl, —CH₂—C₃₋₇cycloalkyl or —COOH; R^(5b) is selected from H, —OH, —OC(O)R^(5ba), —CH₂COOH, —O-benzyl, -pyridyl, or —OC(S)NR^(5bb)R^(5bc); R^(yba) is H, —C₁₋₆alkyl, aryl, —OCH₂-aryl, —CH₂O-aryl, or —NR^(5bb)R^(5bc); R^(5bb) and R^(5ba) are independently selected from H and —C₁₋₄alkyl; R^(5a) is H, —C₁₋₆alkyl, or —C(O)R^(5ca); R^(5ca) is —C₁₋₆alkyl, —C₃₋₇cycloalkyl, aryl, or heteroaryl; R^(5d) is H, —C₁₋₄alkyl, —C₀₋₃alkylenearyl, —NR^(5da)R^(5db), —CH₂SH, or —O—C₁₋₆alkyl; R^(5da) and R^(5db) are independently selected from H and —C₁₋₄alkyl; R^(5e) is H, —C₁₋₆alkyl, —C₁₋₃ alkylenearyl, —C₁₋₃ alkyleneheteroaryl, —C₃₋₇ cyclo alkyl, —CH(CH₃)OC(O)R^(5ca),

R^(5ca) is —O—C₁₋₆alkyl, —O—C₃₋₇cycloalkyl, —NR^(5eb)R^(5ec), or —CH(NH₂)CH₂COOCH₃; R^(5eb) and R^(5ec) are independently selected from H, —C₁₋₄alkyl, and —C₁₋₃alkylenearyl, or are taken together as —(CH₂)₃₋₆₋; R^(5f) is selected from H, —C₀₋₃alkylenearyl, —C₁₋₃alkylene-NR^(5fa)R^(5th), and —C₁₋₃alkylene(aryl)-C₀₋₃alkylene-NR^(5fa)R^(5th); R^(5fa) and R^(5fil) are independently selected from H and —C₁₋₄alkyl; R^(5g) is H, —C₁₋₆alkyl, —C₁₋₃alkylenearyl, or —CH₂—O—(CH₂)₂—OCH₃; R^(5h) is H or —C₁₋₄alkyl; R^(55i) is H, —C₁₋₄alkyl, or —C₀₋₃alkylenearyl; and R^(5j) is —C₁₋₆alkyl, aryl, or —CH₂CH(NH₂)COOH;

R⁶ is selected from —C₁₋₆alkyl, —CH₂—O—(CH₂)₂OCH₃, —C₁₋₆alkylene-OC₁₋₆alkyl, —C₀₋₃alkylenearyl, —C₀₋₃alkyleneheteroaryl, and —C₀₋₃alkylene-C₃₋₇cycloalkyl; and

R⁷ is H or is taken together with R⁶ to form —C₃₋₈cycloalkyl;

wherein: each —CH₂— group in —(CH₂)_(r)— is optionally substituted with 1 or 2 substituents independently selected from —C₁₋₄alkyl and fluoro; each carbon atom in the alkylene moiety in X is optionally substituted with one or more R^(4b) groups and one —CH₂-moiety in X may be replaced with a group selected from —C₃₋₈cycloalkylene-, —CR^(4d)═CH—, and —CH═CR^(4d)—; wherein R^(4b) is selected from —C₀₋₅alkylene-COOR^(4c), —C₁₋₆alkyl, —C₀₋₁alkylene-CONH₂, —C₁₋₂alkylene-OH, —C₀₋₃alkylene-C₃₋₇cycloalkyl, 1H-indol-3-yl, benzyl, and hydroxybenzyl; R^(4c) is H or —C₁₋₄alkyl; and R^(4d) is selected from —CH₂-thiophene and phenyl; each alkyl and each aryl in R¹⁻³, R^(4a-4d), and R₅₋₆ is optionally substituted with 1 to 7 fluoro atoms; each ring in Ar and each aryl and heteroaryl in R¹⁻³ and R⁵⁻⁶ is optionally substituted with 1 to 3 substituents independently selected from —OH, —C₁₋₆alkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl, —CN, halo, —O—C₁₋₆alkyl, —S—C₁₋₆alkyl, —S(O)—C₁₋₆alkyl, —S(O)₂—C₁₋₄alkyl, -phenyl, —NO₂, —NH₂, —NH—C₁₋₆alkyl and —N(C₁₋₆alkyl)₂, wherein each alkyl, alkenyl and alkynyl is optionally substituted with 1 to 5 fluoro atoms; and pharmaceutically acceptable salts thereof.

Another aspect of the invention relates to pharmaceutical compositions comprising a pharmaceutically acceptable carrier and a compound of the invention. Such compositions may optionally contain other therapeutic agents such as diuretics, β₁ adrenergic receptor blockers, calcium channel blockers, angiotensin-converting enzyme inhibitors, AT₁ receptor antagonists, neprilysin inhibitors, non-steroidal anti-inflammatory agents, prostaglandins, anti-lipid agents, anti-diabetic agents, anti-thrombotic agents, renin inhibitors, endothelin receptor antagonists, endothelin converting enzyme inhibitors, aldosterone antagonists, angiotensin-converting enzyme/neprilysin inhibitors, vasopressin receptor antagonists, and combinations thereof. Accordingly, in yet another aspect of the invention, a pharmaceutical composition comprises a compound of the invention, a second therapeutic agent, and a pharmaceutically acceptable carrier. Another aspect of the invention pertains to a combination of active agents, comprising a compound of the invention and a second therapeutic agent. The compound of the invention can be formulated together or separately from the additional agent(s). When formulated separately, a pharmaceutically acceptable carrier may be included with the additional agent(s). Thus, yet another aspect of the invention relates to a combination of pharmaceutical compositions, the combination comprising: a first pharmaceutical composition comprising a compound of the invention and a first pharmaceutically acceptable carrier; and a second pharmaceutical composition comprising a second therapeutic agent and a second pharmaceutically acceptable carrier. The invention also relates to a kit containing such pharmaceutical compositions, for example where the first and second pharmaceutical compositions are separate pharmaceutical compositions.

Compounds of the invention possess both AT₁ receptor antagonist activity and NEP enzyme inhibition activity, and are therefore expected to be useful as therapeutic agents for treating patients suffering from a disease or disorder that is treated by antagonizing the AT₁ receptor and/or inhibiting the NEP enzyme. Thus, one aspect of the invention is directed to a method of treating patients suffering from a disease or disorder that is treated by antagonizing the AT₁ receptor and/or inhibiting the NEP enzyme, comprising administering to a patient a therapeutically effective amount of a compound of the invention. Another aspect of the invention is directed to a method of treating hypertension or heart failure, comprising administering to a patient a therapeutically effective amount of a compound of the invention. Still another aspect of the invention pertains to a method for antagonizing an AT₁ receptor in a mammal comprising administering to the mammal, an AT₁ receptor-antagonizing amount of a compound of the invention. Yet another aspect of the invention pertains to a method for inhibiting a NEP enzyme in a mammal comprising administering to the mammal, a NEP enzyme-inhibiting amount of a compound of the invention.

Compounds of formula I and pharmaceutically acceptable salts thereof, that are of particular interest include those that exhibit an inhibitory constant (pK_(i)) for binding to an AT₁ receptor greater than or equal to about 5.0; in particular those having a plc greater than or equal to about 6.0; in one embodiment those having a pK_(i) greater than or equal to about 7.0; more particularly those having a pK_(i) greater than or equal to about 8.0; and in yet another embodiment, those having a pK_(i) within the range of about 8.0-10.0. Compounds of particular interest also include those having a NEP enzyme inhibitory concentration (plC₅₀) greater than or equal to about 5.0; in one embodiment those having a plC₅₀ greater than or equal to about 6.0; in particular those having a plC₅₀ greater than or equal to about 7.0; and most particularly those having a plC₅₀ within the range of about 7.0-10.0. Compounds of further interest include those having a pK_(i) for binding to an AT₁ receptor greater than or equal to about 7.5 and having a NEP enzyme plC₅₀ greater than or equal to about 7.0.

Since compounds of the invention possess AT₁ receptor antagonist activity and NEP inhibition activity, such compounds are also useful as research tools. Accordingly, one aspect of the invention pertains to a method of using a compound of the invention as a research tool, the method comprising conducting a biological assay using a compound of the invention. Compounds of the invention can also be used to evaluate new chemical compounds. Thus another aspect of the invention relates to a method of evaluating a test compound in a biological assay, comprising: (a) conducting a biological assay with a test compound to provide a first assay value; (b) conducting the biological assay with a compound of the invention to provide a second assay value; wherein step (a) is conducted either before, after or concurrently with step (b); and (c) comparing the first assay value from step (a) with the second assay value from step (b). Exemplary biological assays include an AT₁ receptor binding assay and a NEP enzyme inhibition assay. Still another aspect of the invention is directed to a method of studying a biological system or sample comprising an AT₁ receptor, a NEP enzyme, or both, the method comprising: (a) contacting the biological system or sample with a compound of the invention; and (b) determining the effects caused by the compound on the biological system or sample.

The invention is also directed to processes and intermediates useful for preparing compounds of the invention. Accordingly, another aspect of the invention relates to a process of preparing compounds of the invention comprising the step of coupling a compound of formula 1 with a compound of formula 2:

where: A is —NH₂ and B is —COOH or A is —COOH and B is —NH₂; the sum of a and b is in the range of 0 to 11; Ar* represents Ar—R¹*, where R¹* is R¹ or a protected form of R¹; and R⁵* represents R⁵ or a protected form of R⁵; the carbon atoms in the —(CH₂)_(a) and —(CH₂)_(b) groups may be substituted with one or more R^(4b) groups; and one —CH₂— group in the —(CH₂)_(a) or the —(CH₂)_(b) group may be replaced with —C₃₋₈cycloalkylene-, —CR^(4d)═CH—, or —CH═CR^(4d)—; and optionally deprotecting the product when R¹* is a protected form of R¹ and/or R⁵* is a protected form of R⁵. Another aspect of the invention relates to a process of preparing a pharmaceutically acceptable salt of a compound of formula I, comprising contacting a compound of formula I in free acid or base form with a pharmaceutically acceptable base or acid. In other aspects, the invention is directed to products prepared by any of the processes described herein, as well as novel intermediates used in such process. In one aspect of the invention novel intermediates have formula II, III or IV, as defined herein.

Yet another aspect of the invention is directed to the use of a compound of formula I or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament, especially for the manufacture of a medicament useful for treating hypertension or heart failure. Another aspect of the invention is directed to use of a compound of the invention for antagonizing an AT₁ receptor or for inhibiting a NEP enzyme in a mammal. Still another aspect of the invention pertains to the use of a compound of the invention as a research tool. Other aspects and embodiments of the invention are disclosed herein.

DETAILED DESCRIPTION OF THE INVENTION

The invention is directed to compounds of formula I:

and pharmaceutically acceptable salts thereof.

As used herein, the term “compound of the invention” is intended to include compounds of formula I as well as the species embodied in formulas Ia, Ia-1, Ia-2, Ia-3, Ib, Ib-1, Ib-2, Ib-3, Ic and Id, described below. In addition, the compounds of the invention may also contain several basic or acidic groups (e.g., amino or carboxyl groups) and therefore, such compounds can exist as a free base, free acid, or in various salt forms. All such salt forms are included within the scope of the invention. Furthermore, solvates of compounds of formula I or salts thereof are included within the scope of the invention. Finally, the compounds of the invention may also exist as prodrugs. Accordingly, those skilled in the art will recognize that reference to a compound herein, for example, reference to a “compound of the invention” includes reference to a compound of formula I as well as to pharmaceutically acceptable salts, solvates and prodrugs of that compound unless otherwise indicated. Further, the term “or a pharmaceutically acceptable salt, solvate and/or prodrug thereof” is intended to include all permutations of salts and solvates, such as a solvate of a pharmaceutically acceptable salt.

The compounds of formula I may contain one or more chiral centers and so may exist in a number of stereoisomeric forms. When such chiral centers are present, the invention is directed to racemic mixtures, pure stereoisomers (i.e., enantiomers or diastereomers), stereoisomer-enriched mixtures, and the like unless otherwise indicated. When a chemical structure is depicted without any stereochemistry, it is understood that all possible stereoisomers are encompassed by such structure. Thus, for example, the term “compound of formula I” is intended to include all possible stereoisomers of the compound. Similarly, when a particular stereoisomer is shown or named herein, it will be understood by those skilled in the art that minor amounts of other stereoisomers may be present in the compositions of the invention unless otherwise indicated, provided that the utility of the composition as a whole is not eliminated by the presence of such other isomers. Individual enantiomers may be obtained by numerous methods that are well known in the art, including chiral chromatography using a suitable chiral stationary phase or support, or by chemically converting them into diastereomers, separating the diastereomers by conventional means such as chromatography or recrystallization, then regenerating the original enantiomers. Additionally, where applicable, all cis-trans or E/Z isomers (geometric isomers), tautomeric forms and topoisomeric forms of the compounds of the invention are included within the scope of the invention unless otherwise specified.

Compounds of formula I may contain one or more chiral centers. One possible chiral center could be present in the X portion of the compound. For example, a chiral center exists at a carbon atom in the alkylene moiety in X that is substituted with an Rob group such as —C₁₋₆alkyl, for example —CH₃. This chiral center is present at the carbon atom indicated by the symbol * in the following partial formula:

Another possible chiral center could be present in the —CR⁵R⁶R⁷ portion of the compound, when R⁶ is a group such as —C₁₋₆alkyl, for example —CH₂CH(CH₃)₂, and R⁷ is H. This chiral center is present at the carbon atom indicated by the symbol ** in the following formula:

In one embodiment of the invention, the carbon atom identified by the symbol * and/or ** has the (R) configuration. In this embodiment, compounds of formula I have the (R) configuration at the carbon atom identified by the symbol * and/or ** or are enriched in a stereoisomeric form having the (R) configuration at this carbon atom (or atoms). In another embodiment, the carbon atom identified by the symbol * and/or ** has the (S) configuration. In this embodiment, compounds of formula I have the (S) configuration at the carbon atom identified by the symbol * and/or ** or are enriched in a stereoisomeric form having the (S) configuration at this carbon atom. It is understood that a compound may have a chiral center at both the * and the ** carbon atoms. In such cases, four possible diastereomers can exist. In some cases, in order to optimize the therapeutic activity of the compounds of the invention, for example, as hypertensive agents, it may be desirable that the carbon atom identified by the symbol * and/or ** have a particular (R) or (S) configuration.

The compounds of the invention, as well as those compounds used in their synthesis, may also include isotopically-labeled compounds, that is, where one or more atoms have been enriched with atoms having an atomic mass different from the atomic mass predominately found in nature. Examples of isotopes that may be incorporated into the compounds of formula I, for example, include, but are not limited to, ²H, 3H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, ³⁵S, ³⁶Cl, and ¹⁸F.

The compounds of formula I have been found to possess AT₁ receptor antagonizing activity and NEP enzyme inhibition activity. Among other properties, such compounds are expected to be useful as therapeutic agents for treating diseases such as hypertension. By combining dual activity into a single compound, double therapy can be achieved, that is, AT₁ receptor antagonist activity and NEP enzyme inhibition activity can be obtained using a single active component. Since pharmaceutical compositions containing one active component are typically easier to formulate than compositions containing two active components, such single-component compositions provide a significant advantage over compositions containing two active components. In addition, certain compounds of the invention have also been found to be selective for inhibition of the AT₁ receptor over the angiotensin II type 2 (AT₂) receptor, a property that may have therapeutic advantages.

The nomenclature used herein to name the compounds of the invention is illustrated in the Examples herein. This nomenclature has been derived using the commercially available AutoNom software (MDL, San Leandro, Calif.).

REPRESENTATIVE EMBODIMENTS

The following substituents and values are intended to provide representative examples of various aspects and embodiments of the invention. These representative values are intended to further define and illustrate such aspects and embodiments and are not intended to exclude other embodiments or to limit the scope of the invention. In this regard, the representation that a particular value or substituent is preferred is not intended in any way to exclude other values or substituents from the invention unless specifically indicated.

The invention is directed to compounds of formula I:

The values for r are 0, 1 or 2. In one embodiment, r is 1. Each —CH₂— group in the —(CH₂)_(r)— group may be substituted with 1 or 2 substituents independently selected from —C₁₋₄alkyl (for example, —CH₃) and fluoro. In one particular embodiment, the —(CH₂)_(r)— group is unsubstituted; in another embodiment, one or two —CH₂— groups in —(CH₂)_(r)— are substituted with a —C₁₋₄alkyl group.

Ar represents an aryl group selected from:

Each ring in the Ar moiety may be substituted with 1 to 3 substituents independently selected from —OH, —C₁₋₆alkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl, —CN, halo, —O—C₁₋₆alkyl, —S—C₁₋₆ alkyl, —S(O)—C₁₋₆ alkyl, —S(O)₂—C₁₋₄ alkyl, -phenyl, —NO₂, —NH₂, —NH—C₁₋₆ alkyl and —N(C₁₋₆alkyl)₂. Furthermore, each of the aforementioned alkyl, alkenyl and alkynyl groups are optionally substituted with 1 to 5 fluoro atoms.

In one particular embodiment, each ring in the Ar moiety may be substituted with 1 to 2 substituents independently selected from —OH, —C₁₋₄alkyl (for example, —CH₃), halo (for example bromo, fluoro, chloro, and di-fluoro), —O—C₁₋₄alkyl (for example, —OCH₃), and -phenyl. Exemplary substituted Ar moieties include:

Of particular interest is where Ar is substituted with 1 or 2 halo atoms, particularly fluoro or chloro atoms.

It is understood that the Ar structure:

represents:

In one particular embodiment, Ar is an aryl group selected from:

where Ar is optionally substituted with one fluoro, chloro or bromo atom.

R¹ is selected from —COOR^(1a), —NHSO₂R^(1b), —SO₂NHR^(1d), —SO₂OH, —C(O)NH—SO₂R^(1c), —P(O)(OH)₂, —CN, —OCH(R^(1e))—COOH, tetrazol-5-yl,

The R^(1a) moiety is H, —C₁₋₆alkyl, —C₁₋₃alkylenearyl, —C₁₋₃alkyleneheteroaryl, —C₃₋₇cycloalkyl, —CH(C₁₋₄alkyl)OC(O)R^(1aa), —C₀₋₆alkylenemorpholine,

R^(1aa) is —O—C₁₋₆alkyl, —O—C₃₋₇cycloalkyl, —NR^(1ab)R^(1ac), or —CH(NH₂)CH₂COOCH₃, R^(1ab) and R^(1ac) are independently selected from H, —C₁₋₆alkyl, and benzyl, or are taken together to form —(CH₂)₃₋₆—.

The R^(1b) moiety is R^(1c) or —NHC(O)R^(1c). The R^(1c) group is —C₁₋₆alkyl, —C₀₋₆alkylene-O—R^(1ca), —C₁₋₅alkylene-NR^(1cb)R^(1cc), or —C₀₋₄alkylenearyl. The R^(1ca) moiety is H, —C₁₋₆alkyl, or —C₁₋₆alkylene-O—C₁₋₆alkyl. The R^(1cb) and R^(1cc) groups are independently selected from H and —C₁₋₆alkyl, or are taken together as —(CH₂)₂—O—(CH₂)₂— or —(CH₂)₂—N[C(O)CH₃]—(CH₂)₂—. The R^(1d) moiety is H, R^(1c), —C(O)R^(1c), or —C(O)NHR^(1c). The R^(1e) group is —C₁₋₄alkyl or aryl.

In one embodiment, R¹ is —COOR^(1a) and R^(1a) is H. In another embodiment, R¹ is —COOR^(1a) and R^(1a) is —C₁₋₆alkyl, examples of which include —CH₃, —CH₂CH₃, —(CH₂)₂CH₃, —(CH₂)₂—CF₃, —CH(CH₃)₂, —CH(CH₂F)₂, —CH(CH₃)CF₃, —CH₂CH(CH₃)₂, —C(CH₃)₃, —(CH₂)₃CH₃, and —(CH₂)₂—CF₂CF₃. Thus, examples of R¹ include —C(O)OCH₃, —C(O)OCH₂CH₃, —C(O)O—(CH₂)₂CH₃, —C(O)O—CH₂CH(CH₃)₂, —C(O)OC(CH₃)₃, —C(O)O—(CH₂)₃CH₃, and so forth.

In one embodiment, R¹ is —COOR^(1a) and R^(1a) is —C₁₋₃alkylenearyl, for example, a benzyl group, which may be substituted, examples of which include chlorobenzyl such as 3-chlorobenzyl and 4-chlorobenzyl, fluorobenzyl such as 3-fluorobenzyl, difluorobenzyl such as 2,6-difluorobenzyl, methylbenzyl such as 4-methylbenzyl, methoxybenzyl such as 3-methoxybenzyl, -benzyl-CF₃, and -benzyl-O—CF₃. Thus, examples of R¹ include —C(O)OCH₂-benzyl,

In one embodiment, R¹ is —COOR^(1a) and R^(1a) is —C₁₋₃alkyleneheteroaryl, examples of which include —CH₂-pyridinyl. In one embodiment, R¹ is —COOR^(1a) and R^(1a) is —C₃₋₇cycloalkyl, examples of which include cyclopentyl.

In yet another embodiment R¹ is —COOR^(1a) and R^(1a) is —CH(C₁₋₄alkyl)OC(O)R^(1aa), where R^(1aa) is —O—C₁₋₆alkyl, —O—C₃₋₇cycloalkyl, —NR^(1ab)R^(1ac), or —CH(NH₂)CH₂COOCH₃. R^(1ab) and R^(1ac) are independently selected from H, —C₁₋₆alkyl, and benzyl, or are taken together to form —(CH₂)₃₋₆—. Examples of —O—C₁₋₆alkyl groups include —O—CH₂CH₃ and —O—CH(CH₃)₂. Exemplary —O—C₃₋₇cycloalkyl groups include —O-cyclohexyl. Thus, examples of R¹ include —C(O)OCH(CH₃)OC(O)—O—CH₂CH₃, —C(O)OCH(CH₃)OC(O)—O—CH(CH₃)₂, and —C(O)OCH(CH₃)OC(O)—O-cyclohexyl.

In one embodiment, R¹ is —COOR^(1a) and R^(1a) is —C₀₋₆alkylenemorpholine, examples of which include —(CH₂)₂-morpholine and —(CH₂)₃-morpholine, thus one example of R¹ is —C(O)O(CH₂)₃-morpholin-4-yl. In another embodiment, R^(1a) is

For example, R¹ may be:

In one embodiment, R¹ is —NHSO₂R^(1b) and R^(1b) is R^(1c). The R^(1c) group can be —C₁₋₆alkyl, —C₀₋₆alkylene-O—R^(1ca), —C₁₋₅alkylene-NR^(1cb)R^(1cc), —C₀₋₄alkylenearyl or —C₀₋₄alkyleneheteroaryl. The R^(1ac) moiety is H, —C₁₋₆ alkyl, or —C₁₋₆ alkylene-O—C₁₋₆ alkyl. The R^(1cb) and R^(1cc) groups are independently selected from H and —C₁₋₆alkyl, or are taken together as —(CH₂)₂—O—(CH₂)₂— or —(CH₂)₂—N[C(O)CH₃]—(CH₂)₂—. In one embodiment, R^(1c) is —C₁₋₆alkyl, such that exemplary R¹ groups include —NHSO₂—CH₃ and the fluoro-substituted group, —NHSO₂—CF₃. In another embodiment, R^(1c) is —C₀₋₄alkylenearyl, such that exemplary R¹ groups include —NHSO₂-phenyl. In another embodiment, R^(1c) is —C₀₋₄alkyleneheteroaryl, such that exemplary R¹ groups include —NHSO₂-4,5-dimethylisoxazol-3-yl.

In another embodiment, R¹ is —NHSO₂R^(1b) and R^(1b) is —NHC(O)R^(1c), where R^(1c) is defined above. In a particular embodiment, R¹ is —NHSO₂R^(1b), R^(1b) is —NHC(O)R^(1c), and R^(1c) is —C₁₋₆alkyl or —C₀₋₄alkylenearyl.

In one embodiment, R¹ is —SO₂NHR^(1d) and R^(1d) is H. In another embodiment, R¹ is —SO₂NHR^(1d) and R^(1d) is R^(1c), R^(1c) is defined above. In a particular embodiment, R^(1c) is —C₁₋₆alkyl or —C₀₋₄alkylenearyl. When R^(1c) is —C₁₋₆alkyl, exemplary R¹ groups include the fluoro-substituted groups —SO₂NH—CF₃, —SO₂NH—CHF₂, —SO₂NH—CF₂CH₂F and —SO₂NH—CF₂CF₂CF₃.

In another embodiment, R¹ is —SO₂NHR^(1d) and R^(1d) is —C(O)R^(1c), where R^(1c) is defined above. In one embodiment of particular interest, R^(1c) is —C₁₋₆alkyl or —C₀₋₄alkylenearyl. When R^(1c) is —C₁₋₆alkyl, exemplary R¹ groups include —SO₂NHC(O)CH₃ and —SO₂NHC(O)—(CH₂)₂CH₃. When R^(1c) is —C₀₋₆alkylene-O—R^(1ca) and R^(1ca) is H, exemplary R¹ groups include —SO₂NHC(O)CH₂OH, —SO₂NHC(O)CH(CH₃)OH, and —SO₂NHC(O)C(CH₃)₂OH. When R^(1c) is —C₀₋₆alkylene-O—R^(1ca) and R^(1ca) is —C₁₋₆alkyl, exemplary R¹ groups include —SO₂NHC(O)CH₂—O—CH₃, —SO₂NHC(O)—O—CH₃, and —SO₂NHC(O)—O—CH₂CH₃. When R^(1c) is —C₀₋₆alkylene-O—R^(1ca) and R^(1ca) is —C₁₋₆alkylene-O—C₁₋₆alkyl, exemplary R¹ groups include —SO₂NHC(O)CH₂—O—(CH₂)₂—O—CH₃. When R^(1c) is —C₁₋₅alkylene-NR^(1cb)R^(1cc), exemplary R¹ groups include —SO₂NHC(O)CH₂N(CH₃)₂, —SO₂NHC(O)CH₂NH₂, and —SO₂NHC(O)—CH(CH₃)(NH₂). Another example when R^(1c) is —C₁₋₅ alkylene-NR^(1cb)R^(1cc) is where the R^(1cb) and R^(1cc) are taken together as —(CH₂)₂—O—(CH₂)₂— or —(CH₂)₂—N[C(O)CH₃]-(CH₂)₂—. Such exemplary R¹ groups include:

When R^(1c) is —C₀₋₄alkylenearyl, exemplary R¹ groups include —SO₂NHC(O)-phenyl.

In another embodiment, R¹ is —SO₂NHR^(1d) and R^(1d) is —C(O)NHR^(1c), where R^(1c) is defined above. In a particular embodiment, R^(1c) is —C₁₋₆alkyl or —C₀₋₄alkylenearyl. When R^(1c) is —C₁₋₆alkyl, exemplary R¹ groups include —SO₂NHC(O)NH—CH₂CH₃ and —SO₂NHC(O)NH—(CH₂)₂CH₃. When R^(1c) is —C₀₋₄alkylenearyl, exemplary R¹ groups include —SO₂NHC(O)NH-phenyl.

In another embodiment, R¹ is —SO₂OH, and in still another embodiment, R¹ is —P(O)(OH)₂. In yet another embodiment, R¹ is —CN.

In another embodiment, R¹ is —C(O)NH—SO₂R^(1c), where R^(1c) is defined above. In a particular embodiment, R^(1c) is —C₁₋₆alkyl or —C₀₋₄alkylenearyl. When R^(1c) is —C₁₋₆alkyl, exemplary R¹ groups include —C(O)NH—SO₂—CH₃, —C(O)NH—SO₂—CH₂CH₃ and the fluoro-substituted —C(O)NH—SO₂—CF₃ group.

In another embodiment, R¹ is —O—CH(R^(1e))—COOH, where R^(1e) is —C₁₋₄alkyl or aryl.

Examples of such R¹ groups include, —O—CH(CH₃)—COOH and —O—CH(phenyl)-COOH.

In an embodiment of particular interest, R¹ is tetrazol-5-yl. In another embodiment, R¹ is:

Each alkyl and each aryl in R¹ is optionally substituted with 1 to 7 fluoro atoms. In addition, the term “alkyl” is intended to include divalent alkylene groups such as those present in —C₁₋₃ alkylenearyl and —C₁₋₃ alkyleneheteroaryl, for example.

Further, each aryl and heteroaryl group that might be present in R¹ may be substituted with 1 to 3-OH, —C₁₋₆alkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl, —CN, halo, —O—C₁₋₆alkyl, —S—C₁₋₆alkyl, —S(O)—C₁₋₆alkyl, —S(O)₂—C₁₋₄alkyl, -phenyl, —NO₂, —NH₂, —NH—C₁₋₆alkyl, or —N(C₁₋₆alkyl)₂ groups. Further, each of the aforementioned alkyl, alkenyl and alkynyl groups may be substituted with 1 to 5 fluoro atoms. It is understood that when referring to “each alkyl,” “each aryl” and “each heteroaryl” in R¹, the term also includes any alkyl, aryl, and heteroaryl groups that might be present in the R^(1a) through R^(1e) moieties.

The values for n are 0, 1, 2 or 3. In one embodiment, n is 0. In another embodiment, n is 1, and in one particular embodiment, R² is at the 4 position of the benzoimidazole ring, as depicted below:

Each R² is independently selected from halo, —NO₂, —C₁₋₆alkyl, —C₂₋₆alkenyl, —C₃₋₆cycloalkyl, —CN, —C(O)R^(2a), —C₀₋₅alkylene-OR^(2b), —C₀₋₅alkylene-NR^(2c)R^(2d), —C₀₋₃alkylenearyl, and —C₀₋₃alkyleneheteroaryl. The R^(2a) moiety is H, —C₁₋₆alkyl, —C₃₋₆cycloalkyl, —OR^(2b), or —NR^(2c)R^(2d). R^(2b) is selected from H, —C₁₋₆alkyl, —C₃₋₆cycloalkyl, and —C₀₋₁alkylenearyl; and R^(2a) and R^(2d) are independently selected from H, —C₁₋₄alkyl, and —C₀₋₁alkylenearyl.

In one particular embodiment, R² is halo, for example, chloro. In yet another embodiment, R² is —C₁₋₆alkyl such as —CH₃, and fluoro-substituted alkyl groups such as —CH₂F and —CF₃. In another embodiment R² is —C₀₋₅alkylene-OR^(2b) and R^(2b) is H; one such R² group is —CH₂OH.

Each alkyl and each aryl in R² is optionally substituted with 1 to 7 fluoro atoms. It is understood that when referring to the “alkyl” in R², the term includes any alkyl groups that might be present in the R^(2a), R^(2b), R^(2a) and R^(2d) moieties. In addition, each aryl and heteroaryl in R², for example in —C₀₋₃alkylenearyl and —C₀₋₃alkyleneheteroaryl, may be substituted with 1 to 3-OH, —C₁₋₆alkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl, —CN, halo, —O—C₁₋₆alkyl, —S—C₁₋₆ alkyl, —S(O)—C₁₋₆ alkyl, —S(O)₂—C₁₋₄ alkyl, -phenyl, —NO₂, —NH₂, —NH—C₁₋₆ alkyl, or —N(C₁₋₆alkyl)₂ groups. Further, each of the aforementioned alkyl, alkenyl and alkynyl groups may be substituted with 1 to 5 fluoro atoms. It is understood that when referring to the “aryl” or the “heteroaryl” in R², the term includes any aryl groups that might be present in the R^(2a), R^(2b), R^(2a) and R^(2d) moieties.

R³ is selected from —C₁₋₁₀alkyl, —C₂₋₁₀ alkenyl, —C₃₋₁₀alkynyl, —C₀₋₃alkylene-C₃₋₇cycloalkyl, —C₂₋₃alkenylene-C₃₋₇cycloalkyl, —C₂₋₃alkynylene-C₃₋₇cycloalkyl, —C₀₋₅alkylene-NR^(3a)—C₀₋₅alkylene-R^(3b), —C₀₋₅alkylene-O—C₀₋₅alkylene-R^(3b), —C₀₋₅alkylene-S—C₁₋₅alkylene-R^(3b), and —C₀₋₃alkylenearyl. R^(1a) can be H, —C₁₋₆alkyl, —C₃₋₇cycloalkyl, or —C₀₋₃alkylenearyl (for example, —C₀₋₁alkylenearyl such as phenyl and benzyl). R^(3b) can be H, —C₁₋₆alkyl, —C₃₋₇cycloalkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl, or aryl (such as phenyl).

In addition, each alkyl and each aryl in R³ is optionally substituted with 1 to 7 fluoro atoms, where the term “alkyl” is intended to include divalent alkylene groups such as those present in —C₀₋₃alkylene-C₃₋₇cycloalkyl and —C₀₋₃alkylenearyl, for example. Each aryl in R³, for example in —C₀₋₃alkylenearyl or phenyl, may be substituted with 1 to 3-OH, —C₁₋₆alkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl, —CN, halo, —O—C₁₋₆alkyl, —S—C₁₋₆alkyl —S(O)—C₁₋₆alkyl, —S(O)₂—C₁₋₄alkyl, -phenyl, —NO₂, —NH₂, —NH—C₁₋₆alkyl, or —N(C₁₋₆alkyl)₂ groups. Further, each of the aforementioned alkyl, alkenyl and alkynyl groups may be substituted with 1 to 5 fluoro atoms. It is understood that when referring to “each alkyl” or “each aryl” in R³, the term includes any alkyl and aryl groups that might be present in the R^(3a) and R^(3b) moieties.

In one embodiment, R³ is —C₁₋₁₀alkyl, examples of which include, —CH₂CH₃, —(CH₂)₂CH₃, and —(CH₂)₃CH₃. In another embodiment, R³ is —C₂₋₇alkyl; and in yet another embodiment, R³ is —C₂₋₅alkyl, for example, —(CH₂)₂CH₃.

In another embodiment, R³ is —C₂₋₁₀alkenyl such as —CH₂CH═CHCH₃. In yet another embodiment, R³ is —C₃₋₁₀alkynyl such as —CH₂CCCH₃.

In another embodiment, R³ is —C₀₋₃alkylene-C₃₋₇cycloalkyl such as -cyclopropyl, —CH₂-cyclopropyl, cyclopentyl, —CH₂-cyclopentyl, —(CH₂)₂-cyclopentyl, and —CH₂-cyclohexyl. In a particular embodiment, R³ is —C₀₋₁alkylene-C₃₋₅cycloalkyl. In one embodiment, R³ is —C₂₋₃alkenylene-C₃₋₇cycloalkyl such as —CH₂CH═CH-cyclopentyl; and in another embodiment, R³ is —C₂₋₃alkynylene-C₃₋₇cycloalkyl such as —CH₂C≡C-cyclopentyl.

In yet another embodiment, R³ is —C₀₋₅alkylene-NR^(3a)—C₀₋₅alkylene-R^(3b). In one particular embodiment, R^(3a) is H and R^(3b) is —C₁₋₆alkyl. Examples of such R³ groups include —NHCH₂CH₃, —NHCH(CH₃)₂, —NH(CH₂)₂CH₃, —NH(CH₂)₃CH₃, —NHCH(CH₃)CH₂CH₃, —NH(CH₂)₄—CH₃, and —NH(CH₂)₅CH₃.

In one embodiment, R³ is —C₀₋₅alkylene-O—C₀₋₅alkylene-R^(3b). In one particular embodiment, R^(3b) is selected from H, —C₁₋₆alkyl and aryl. Examples of such R³ groups include —OCH₃, —OCH₂CH₃, —OCH(CH₃)₂, —O(CH₂)₂CH₃, —O(CH₂)₃CH₃, —OCH₂CH(CH₃)₂, —O-phenyl, and —O-benzyl. In another embodiment, R³ is —C₀₋₅alkylene-O—C₀₋₅alkylene-R^(3b), where R^(3b) is —C₁₋₆alkyl, and in another embodiment, R³ is —O—C₁₋₅alkyl.

In another embodiment, R³ is —C₀₋₅alkylene-S—C₁₋₅alkylene-R^(3b), and in one particular embodiment R^(3b) is H, such as when R³ is —CH₂—S—CH₂CH₃. In another embodiment, R³ is —C₀₋₃alkylenearyl, such as phenyl, benzyl, and —(CH₂)₂-phenyl.

X is —C₁₋₁₂alkylene-, where at least one —CH₂— moiety in the alkylene is replaced with a —NR^(4a)—C(O)— or —C(O)—NR^(4a)— moiety. Thus X is selected from —C₁alkylene-, —C₁alkylene-, —C₂alkylene-, —C₃alkylene-, —C₄alkylene-, —C₅alkylene-, —C₆alkylene-, —C₇alkylene-, —C₈alkylene, —C₉alkylene-, —C₁₀alkylene-, —C₁₁alkylene-, and —C₁₂alkylene-. R^(4a) is selected from H, —OH, and —C₁₋₄alkyl. In one embodiment, R^(4a) is H.

Each carbon atom in this C₁₋₁₂alkylene moiety may be substituted with one or more R^(4b) groups. R^(4b) is selected from —C₀₋₅alkylene-COOR^(4c), —C₁₋₆alkyl, —C₀₋₁alkylene-CONH₂, —C₁₋₂alkylene-OH, —C₀₋₃alkylene-C₃₋₇cycloalkyl, 1H-indol-3-yl, benzyl, and hydroxybenzyl, where R^(4a) is H or —C₁₋₄alkyl. In one embodiment, the carbon atoms in —C₁₋₁₂alkylene- are unsubstituted with R^(4b) groups. In another embodiment, 1 or 2 carbon atoms are substituted with one or two R^(4b) groups; and in one such embodiment, R^(4b) is —COOH, benzyl or —C₁₋₆alkyl, including —C₁₋₃alkyl groups such as —CH₃ and —CH(CH₃)₂.

In addition, one —CH₂— moiety in X may be replaced with a group selected from —C₃₋₈cycloalkylene- (for example, cyclohexylene), —CR^(4d)═CH—, and —CH═CR^(4d)—, where R^(4d) is selected from —CH₂-thiophene and phenyl. In one embodiment, none of the —CH₂-moieties are so replaced.

Each alkyl and each aryl in R^(4a), R^(4b), R^(4a), and R^(4d) may be substituted with 1 to 7 fluoro atoms, and the term “alkyl” is intended to include divalent alkylene groups such as that present in —C₀₋₅alkylene-COOR^(4c), for example. It is noted that the R^(4b) group, —C₀₋₃alkylene-C₃₋₇cycloalkyl, is intended to include a C₃₋₇cycloalkyl linked to the X —C₁₋₁₂alkylene- chain by a bond as well as a C₃₋₇cycloalkyl that is directly attached to the chain, as illustrated below:

In one embodiment, one to four —CH₂— moieties are replaced with —NR^(4a)—C(O)— or —C(O)—NR^(4a)— moieties; in another embodiment one —CH₂— moiety is replaced, examples of which include: —C(O)NH—, —NHC(O)—, and —CH₂—NHC(O)—. In one embodiment, X is C₁₋₆alkylene and 1, 2, 3, or 4—CH₂— moieties are replaced with a —NR^(4a)—C(O)— or —C(O)—NR^(4a)— moiety; in another embodiment X is —C₁₋₃alkylene- and 1 or 2—CH₂— moieties are replaced. In one embodiment X is —C₁₋₂alkylene- and one —CH₂— moiety is replaced. When more than one —CH₂— moiety in —C₁₋₁₂alkylene- is replaced with a —NR^(4a)—C(O)— or —C(O)—NR^(4a)— moiety, the replaced moieties may be contiguous or non-contiguous.

Exemplary X groups include the following, which depict, examples where one or more —CH₂— moieties are replaced with —NR^(4a)—C(O)— or —C(O)—NR^(4a)— moieties, examples where —CH₂— moieties are replaced with a group selected from —C₃₋₈cycloalkylene-, —CR^(4d)═CH—, and —CH═CR^(4d)—, as well as examples where carbon atoms in the —C₁₋₁₂alkylene- group are substituted with one or more R^(ob) groups:

—C₁alkylene- with one —CH₂— moiety replaced:

—C(O)NH—

—NHC(O)—

—C₂alkylene- with one —CH₂— moiety replaced:

—CH₂—NHC(O)—

—C(O)NH—CH₂—

—CH₂—C(O)NH—

—CH[CH(CH₃)₂]—C(O)NH—

—C₂alkylene- with two —CH₂— moieties replaced:

—C(O)NH—NHC(O)—

—CH═C(—CH₂-2-thiophene)-C(O)NH—

—C₃alkylene- with one —CH₂— moiety replaced:

—(CH₂)₂—NHC(O)—

—CH(CH₃)—CH₂—NHC(O)—

—CH[CH(CH₃)₂]—CH₂—NHC(O)—

—CH(COOH)—CH₂—NHC(O)—

—CH₂—CH(COOH)—NHC(O)—

—C₃alkylene- with two —CH₂— moieties replaced:

—NHC(O)—CH₂—NHC(O)—

—C₄alkylene- with one —CH₂— moiety replaced:

—(CH₂)₃—NHC(O)—

—C(O)NH—CH₂—CH(COOH)—CH₂—

—C₄alkylene- with two —CH₂— moieties replaced:

—C(O)NH—CH(benzyl)-CH₂—NHC(O)—

—C(O)NH—CH(benzyl)-CH₂—C(O)NH—

—CH₂—NHC(O)—CH₂—NHC(O)—

—C₄alkylene- with three —CH₂— moieties replaced:

—CH₂—NHC(O)-cyclohexylene-NHC(O)—

—CH₂—N(OH)C(O)-cyclohexylene-NHC(O)—

—C₅alkylene- with two —CH₂— moieties replaced:

—CH₂—NHC(O)—CH₂—CH(COOH)—NHC(O)—

—CH₂—NHC(O)—(CH₂)₂—NHC(O)—

—C(O)NH—(CH₂)₂—C(O)N(OH)—CH₂—

—C(O)NH—(CH₂)₂—CH(COOH)—NHC(O)—

—CH(COOH)—CH₂—NHC(O)—CH₂—NHC(O)—

—(CH₂)₂—NHC(O)-cyclohexylene-NHC(O)—

—C₆alkylene- with two —CH₂— moieties replaced:

—C(O)NH—(CH₂)₄—NHC(O)—

—CH₂—NHC(O)—(CH₂)₂—CH(COOH)—NHC(O)—

—C(O)NH—(CH₂)₃—CH(COOH)—NHC(O)—

—C₆alkylene- with three —CH₂— moieties replaced:

—C(O)NH—(CH₂)₂—NHC(O)—CH₂—NHC(O)—

—C₆alkylene- with four —CH₂— moieties replaced:

—C(O)NH—(CH₂)₂—NHC(O)-cyclohexylene-NHC(O)—

—C₇alkylene- with two —CH₂— moieties replaced:

—CH₂—NHC(O)—(CH₂)₄—NHC(O)—

—C(O)NH—(CH₂)₄—CH(COOH)—NHC(O)—

—C₇alkylene- with three —CH₂— moieties replaced:

—CH[CH(CH₃)₂]—C(O)NH—(CH₂)₂—NHC(O)—CH₂—NHC(O)—

—C₇alkylene- with four —CH₂— moieties replaced:

—CH₂—NHC(O)—(CH₂)₂—NHC(O)-cyclohexylene-NHC(O)—

—CH₂—C(O)NH—(CH₂)₂—NHC(O)-cyclohexylene-NHC(O)—

—C₈alkylene- with three —CH₂— moieties replaced:

—C(O)NH—(CH₂)₄—NHC(O)—CH₂—NHC(O)—

—C₈alkylene- with four —CH₂— moieties replaced:

—C(O)NH—(CH₂)₄—NHC(O)-cyclohexylene-NHC(O)—

—C₉alkylene- with two —CH₂— moieties replaced:

—CH₂—NHC(O)—(CH₂)₆—NHC(O)—

—C₉alkylene- with four —CH₂— moieties replaced:

—CH₂—NHC(O)—(CH₂)₄—NHC(O)-cyclohexylene-NHC(O)—

—C₉alkylene- with four —CH₂— moieties replaced:

—C(O)NH—(CH₂)₆—NHC(O)-cyclohexylene-NHC(O)—

—C₁₁alkylene- with three —CH₂— moieties replaced:

—CH(CH(CH₃)₂)—C(O)NH—(CH₂)₆—NHC(O)—CH₂—NHC(O)—

—C₁₁alkylene- with four —CH₂— moieties replaced:

—CH₂—NHC(O)—(CH₂)₆—NHC(O)-cyclohexylene-NHC(O)—

In one embodiment, X is selected from —C(O)NH—, —NHC(O)—, and —CH₂—NHC(O)—.

R⁵ is selected from —C₀₋₃alkylene-SR^(5a), —C₀₋₃alkylene—C(O)NR^(5b)R^(5c), —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d), —NH—C₀₋₁alkylene-P(O)(OR^(5e))₂, —C₀₋₃alkylene-P(O)OR^(5e)R^(5f), —C₀₋₂alkylene-CHR^(5g)—COOH, —C₀₋₃alkylene—C(O)NR^(5l)—CHR^(5l)—COOH, and —C₀₋₃alkylene-S—SR^(5j). Each alkyl and each aryl in R⁵ is optionally substituted with 1 to 7 fluoro atoms, where the term “alkyl” is intended to include divalent alkylene groups such as those present in —C₀₋₃alkylene-SR^(5a) and —C₀₋₃alkylene-P(O)OR^(5e)R^(5f), for example. Each aryl and heteroaryl in R⁵ may be substituted with 1 to 3-OH, —C₁₋₆alkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl, —CN, halo, —O—C₁₋₆alkyl, —S—C₁₋₆alkyl, —S(O)—C₁₋₆alkyl, —S(O)₂—C₁₋₄alkyl, -phenyl, —NO₂, —NH₂, —NH—C₁₋₆alkyl, or —N(C₁₋₆alkyl)₂ groups. Further, each of the aforementioned alkyl, alkenyl and alkynyl groups may be substituted with 1 to 5 fluoro atoms. It is understood that when referring to “each alkyl”, “each aryl”, and “each heteroaryl” in R⁵, the terms also R^(5aa), R^(5ab), include any alkyl, aryl, and heteroaryl groups that might be present in the R^(5a-5j), R^(5ba), R^(5bb), R^(5bc), R^(5ca), R^(5da), R^(5db), R^(5ea), R^(5eb), R^(5ec), R^(fa), and R^(5fb) moieties (which are defined below).

In one embodiment, R⁵ is —C₀₋₃alkylene-SR^(5a). R^(5a) is H or —C(O)—R^(5aa). The R^(5aa) group is R^(5aa) is —C₁₋₆alkyl, —C₀₋₆alkylene-C₃₋₇cycloalkyl, -aminoC₄₋₇cycloalkyl, —C₀₋₆alkylenearyl, —C₀₋₆alkyleneheteroaryl, —C₀₋₆alkylenemorpholine, —C₀₋₆alkylenepiperazine—CH₃, —C₀₋₆alkylenepiperidine, —C₀₋₆alkylenepiperidine—CH₃, —CH[N(R^(5ab))₂]-aa where aa is an amino acid side chain, —C₀₋₆alkylene-CH[N(R^(5ab))₂]—R^(5ac), -2-pyrrolidine, -2-tetrahydrofuran, —C₀₋₆alkylene-OR^(5ab), —O—C₀₋₆alkylenearyl, —C₁₋₂alkylene-OC(O)—C₁₋₆alkyl, —C₁₋₂alkylene-OC(O)—C₀₋₆alkylenearyl, —O—C₁₋₂alkylene-OC(O)O—C₁₋₆alkyl, —C₁₋₄alkylene-COOH, or -arylene-COOH. The R^(5ab) group is H or —C₁₋₆alkyl, and the R^(5ac) group is H, —C₁₋₆alkyl, —CH₂—C₃₋₇cycloalkyl or —COOH. In one specific embodiment, R^(5a) is H, for example, R⁵ may be —SH or —CH₂SH. In another embodiment, R^(5a) is —C(O)—R^(5aa), where R^(5aa) is —C₁₋₆alkyl. Exemplary —C₁₋₆alkyl groups include —CH₃, —CH₂CH₃, —CH(CH₃)₂, —(CH₂)₂CH₃, —C(CH₃)₃, and —CH₂CH(CH₃)₂. Thus, examples of R⁵ include —SC(O)CH₃, —CH₂SC(O)CH₃, —CH₂SC(O)CH₂CH₃, —CH₂SC(O)CH(CH₃)₂, —CH₂SC(O)(CH₂)₂CH₃, and —CH₂SC(O)C(CH₃)₃, and —CH₂SC(O)CH₂CH(CH₃)₂. In one embodiment, R^(5a) is selected from H and —C(O)—C₁₋₆alkyl.

In another embodiment, R^(5aa) is —C(O)—R^(5aa), where R^(5aa) is —C₀₋₆alkylene-C₃₋₇cycloalkyl. Exemplary C₃₋₇cycloalkyl groups include cyclopentyl and cyclohexyl. Thus, examples of R⁵ include —CH₂SC(O)-cyclopentyl, —CH₂SC(O)-cyclohexyl, and —CH₂SC(O)—CH₂-cyclopentyl.

In another embodiment, R^(5a) is —C(O)—R^(5aa), where R^(5aa) is -aminoC₄₋₇cycloalkyl. Such examples of R⁵ include —CH₂SC(O)-aminocyclobutane, —CH₂SC(O)-aminocyclopentane, and —CH₂SC(O)-aminocyclohexane, depicted as:

In another embodiment, R^(5a) is —C(O)—R^(5aa), where R^(5aa) is —C₀₋₆alkylenearyl and the aryl is optionally substituted with 1 to 3 substituents such as —O—C₁₋₆alkyl and —NH₂. Exemplary aryl groups include phenyl and -phenyl-OCH₃. Thus, examples of R⁵ include —CH₂SC(O)-phenyl and —CH₂SC(O)-benzyl and with substituted aryl groups such as —CH₂SC(O)-phenyl-OCH₃, —CH₂SC(O)-2-aminophenyl, —CH₂SC(O)-3-aminophenyl, and —CH₂SC(O)-4-aminophenyl.

In yet another embodiment, R^(5a) is —C(O)—R^(5aa), where R^(5aa) is —C₀₋₆alkyleneheteroaryl. Exemplary heteroaryl groups include furanyl, thienyl and pyridinyl. Thus, examples of R⁵ include: —CH₂SC(O)-2-pyridine, —CH₂SC(O)-3-pyridine, and —CH₂ SC(O)-4-pyridine.

In another embodiment, R^(5a) is —C(O)—R^(5aa), where R^(5aa) is —C₀₋₆alkylenemorpholine:

more particularly, —C₁₋₃alkylenemorpholine. Thus, examples of R⁵ include —CH₂S—C(O)CH₂-morpholin-4-yl, —CH₂S—C(O)(CH₂)₂-morpholin-4-yl, and —CH₂SC(O)(CH₂)₃— morpholin-4-yl. In another embodiment, R^(5a) is —C(O)—R^(5aa), where R^(5aa) is —C₀₋₆alkylenepiperazine—CH₃. Thus, examples of R⁵ include —CH₂SC(O)(CH₂)₂-4-methylpiperazin-1-yl.

In another embodiment, R^(5a) is —C(O)—R^(5aa), where R^(5aa) is —C₀₋₆alkylenepiperidine. Thus, examples of R⁵ include —CH₂SC(O)—CH₂-2-piperidin-1-yl. In another embodiment, R^(5a) is —C(O)—R^(5aa), where R^(5aa) is —C₀₋₆alkylenepiperidine—CH₃. Thus, examples of R⁵ include —CH₂ SC(O)—CH₂-2-methylpiperidin-1-yl, —CH₂SC(O)—CH₂-3-methylpiperidin-1-yl, and —CH₂ SC(O)(CH₂)₂-4-methylpiperazin-1-yl.

In one embodiment, R^(5a) is —C(O)—R^(5aa), where R^(5aa) is —CH[N(R^(5ab))₂]-aa where aa is an amino acid side chain. For example, the amino acid side chain could be —CH(CH₃)₂ (valine side chain), —CH₂CH(CH₃)₂ (leucine side chain), —CH(CH₃)CH₂CH₃ (isoleucine side chain), —CH₂COOH (aspartic acid side chain), —(CH₂)₂COOH (glutamic acid side chain), —CH(OH)(CH₃) (threonine side chain), -benzyl (phenylalanine side chain), -4-hydroxybenzyl (tyrosine side chain), and —(CH₂)₂SCH₃ (methionine side chain). Thus, other examples of R⁵ include —CH₂SC(O)CH(NH₂)—CH(CH₃)₂, —CH₂SC(O)CH(NH₂)—CH₂CH(CH₃)₂, —CH₂SC(O)CH(NH₂)—CH(CH₃)CH₂CH₃, —CH₂SC(O)CH(NH₂)—CH₂COOH, —CH₂SC(O)CH(NH₂)—(CH₂)₂COOH, —CH₂SC(O)CH(NH₂)—CH(OH)(CH₃), —CH₂SC(O)—CH(NH₂)-benzyl, —CH₂ SC(O)CH(NH₂)-4-hydroxybenzyl, —CH₂ SC(O)CH(NH₂)—(CH₂)₂SCH₃.

In one embodiment, R^(5a) is —C(O)—R^(5aa), where R^(5aa) is —C₀₋₆alkylene-CH[N(R^(5ab))₂]—R^(5ac) and R^(5ac) is H. Such examples of R⁵ include —CH₂SC(O)CH₂N(CH₃)₂, —CH₂SC(O)(CH₂)₂N(CH₃)₂, and —CH₂SC(O)(CH₂)₃N(CH₃)₂. In another embodiment, R^(5a) is —C(O)—R^(5aa), where R^(5aa) is —C₀₋₆alkylene-CH[N(R^(5ab))₂]—R^(5ac) and R^(5ac) is —C₁₋₆alkyl. Such examples of R⁵ include —CH₂SC(O)CH(NH₂)—CH₂CH₃, —CH₂SC(O)CH(NH₂)—(CH₂)₂CH₃, —CH₂SC(O)CH(NH₂)—C(CH₃)₃, and —CH₂SC(O)—CH(NH₂)—(CH₂)₃CH₃. In another embodiment, R^(5a) is —C(O)—R^(5aa), where R^(5aa) is —C₀₋₆alkylene-CH[N(R^(5ab))₂]—R^(5ac) and R^(5ac) is —CH₂—C₃₋₇cycloalkyl. Such examples of R⁵ include CH₂SC(O)CH(NH₂)—CH₂-cyclobutyl and —CH₂SC(O)CH(NH₂)—CH₂-cyclohexyl. In yet another embodiment, R^(5a) is —C(O)—R^(5aa), where R^(5aa) is —C₀₋₆alkylene-CH[N(R^(5ab))₂]—R^(5ac) and R^(5ac) is —COOH. Such examples of R⁵ include —CH₂SC(O)CH₂CH(NH₂)COOH and —CH₂SC(O)(CH₂)₂CH(NH₂)—COOH.

In yet another embodiment, R^(5a) is —C(O)—R^(5aa), where R^(5aa) is -2-pyrrolidine or -2-tetrahydrofuran:

Thus, examples of R⁵ include —CH₂SC(O)-2-pyrrolidine and —CH₂SC(O)-tetrahydrofuran-2-yl.

In another embodiment, R^(5a) is —C(O)—R^(5aa), where R^(5aa) is —C₀₋₆alkylene-OR^(5ab). In one embodiment, R^(5ab) is H, such that R^(5a) is —C(O)—C₀₋₆alkylene-OH. In another embodiment, R^(5ab) is —C₁₋₆alkyl, such that R^(5a) is —C(O)—C₀₋₆alkylene-O—C₁₋₆alkyl, for example, R⁵ may be —CH₂SC(O)—O—CH₂CH₃.

In another embodiment, R^(5a) is —C(O)—R^(5aa), where R^(5aa) is —O—C₀₋₆alkylenearyl. In yet another embodiment, R^(5a) is —C(O)—R^(5aa), where R^(5aa) is —C₁₋₂alkylene-OC(O)—C₁₋₆alkyl. One example of such an R⁵ group is —CH₂SC(O)CH₂—OC(O)CH₃. In another embodiment, R^(5a) is —C(O)—R^(5aa), where R^(5aa) is —C₁₋₂alkylene-OC(O)—C₀₋₆alkylenearyl. In yet another embodiment, R^(5a) is —C(O)—R^(5aa), where R^(5aa) is —O—C₁₋₂alkylene-OC(O)O—C₁₋₆alkyl, for example, R⁵ may be —CH₂SC(O)OCH(CH₃)—OC(O)O—CH(CH₃)₂.

In another embodiment, R^(5a) is —C(O)—R^(5aa), where R^(5aa) is —C₁₋₄alkylene-COOH. Thus, examples of R⁵ include —CH₂SC(O)(CH₂)₂COOH. In another embodiment, R^(5a) is —C(O)—R^(5aa), where R^(5aa) is -arylene-COOH. Thus, examples of R⁵ include —CH₂SC(O)-3-carboxyphenyl and —CH₂SC(O)-4-carboxyphenyl.

In one embodiment, R⁵ is —C₀₋₃alkylene—C(O)NR^(5b)R^(5c). The R^(5b) moiety is H, —OH, —OC(O)R^(5ba), —CH₂COOH, —O-benzyl, -pyridyl, or —OC(S)NR^(5bb)R^(5bc). R^(5ba) is H, —C₁₋₆alkyl, aryl, —OCH₂-aryl (for example, —OCH₂-phenyl), —CH₂O-aryl (for example, —CH₂O-phenyl), or —NR^(5bb)R^(5bc). The R^(5bb) and R^(5ba) moieties are independently selected from H and —C₁₋₄alkyl. In one embodiment, R^(5b) is —OH or —OC(O)R^(5ba), where —R^(5ba) is —C₁₋₆alkyl. R^(5a) is H, —C₁₋₆alkyl, or —C(O)R^(5ca), R^(5ca) is —C₁₋₆alkyl, —C₃₋₇cycloalkyl, aryl, or heteroaryl. In one particular embodiment, R^(5c) is H. In another embodiment, R^(5b) is —OH and R^(5c) is H, for example, R⁵ may be —C(O)NH(OH) or —CH₂C(O)NH(OH). In another embodiment, R^(5b) is —OC(O)R^(5ba), where —R^(5ba) is —C₁₋₆alkyl and R^(5a) is H, for example, R⁵ may be —C(O)N[OC(O)CH₃]H or —C(O)N[OC(O)C(CH₃)₃]H. In still another embodiment, both R^(5b) and R^(5c) are H, for example, R⁵ may be —C(O)NH₂. In another embodiment, R^(5b) is —CH₂COOH and R^(5a) is H, for example, R⁵ may be —C(O)NH(CH₂COOH). In yet another embodiment, R^(5b) is —OC(O)R^(5ba), where —R^(5ba) is —O—CH₂-aryl or —CH₂—O-aryl, for example, R⁵ may be —CH₂—C(O)NH[OC(O)OCH₂-phenyl] and —CH₂—C(O)N[OC(O)CH₂O-phenyl]H.

In another embodiment, R^(5b) is —OC(S)NR^(5bb)R^(5bc), where R^(5bb) and R^(5bc) are both —C₁₋₄allyl, for example, R^(5b) may be —O—C(S)N(CH₃)₂. In another embodiment, R^(5b) is —OC(S)NR^(5bb)R^(5bc) and R^(5a) is H, for example, R⁵ may be —CH₂—C(O)N[OC(S)N(CH₃)₂]H.

In one embodiment, R⁵ is —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d). R^(5d) is H, —C₁₋₄alkyl, —C₀₋₃alkylenearyl, —NR^(5da)R^(5db), —CH₂SH, or —O—C₁₋₆alkyl. The R^(5da) and R^(5db) moieties are independently selected from H and —C₁₋₄alkyl. In another embodiment, R^(5b) is —OH and R^(5d) is H, for example, R⁵ may be —CH₂—N(OH)C(O)H. In another embodiment, R^(5b) is —OH and R^(5d) is —C₁₋₄alkyl, for example, R⁵ may be —CH₂—N(OH)C(O)CH₃.

In another embodiment, R^(5b) is H and R^(5d) is —CH₂SH, for example, R⁵ may be —NHC(O)CH₂SH or —CH₂NHC(O)—CH₂SH.

In yet another embodiment, R⁵ is —NH—C₀₋₁alkylene-P(O)(OR^(5e))₂. The R^(5e) moiety is H, —C₁₋₆alkyl, —C₁₋₃alkylenearyl, —C₁₋₃alkyleneheteroaryl, —C₃₋₇cycloalkyl, —CH(CH₃)—O—C(O)R^(5ea),

The R^(5ea) group is —O—C₁₋₆alkyl, —O—C₃₋₇cycloalkyl, —NR^(5eb)R^(5aa), or —CH(NH₂)CH₂COOCH₃. R^(5eb) and R^(5ec) are independently selected from H, —C₁₋₄alkyl, and —C₁₋₃alkylenearyl (for example, benzyl). R^(5eb) and R^(5ec) may also be taken together to form —(CH₂)₃₋₆—. In one embodiment, R^(5e) is H, for example, R⁵ may be —NH—CH₂—P(O)(OH)₂.

In one embodiment, R⁵ is —C₀₋₃alkylene-P(O)OR^(5e)R^(5f). The R^(5f) moiety is H, —C₁₋₄alkyl, —C₀₋₃alkylenearyl, —C₁₋₃alkylene-NR^(5fa)R^(5th), or —C₁₋₃alkylene(aryl)-C₀₋₃alkylene-NR^(5fa)R^(5fb). The R^(5fa) and R^(5fb) groups are independently selected from H and —C₁₋₄alkyl. In one embodiment, R^(5e) is H, for example, R⁵ may be —C₀₋₃alkylene-P(O)(OH)R^(5f).

In one embodiment, R⁵ is —C₀₋₂alkylene-CHR^(5g)—COOH. The R^(5g) moiety is H, —C₁₋₆alkyl, —C₁₋₃alkylenearyl, or —CH₂—O—(CH₂)₂—OCH₃. In one embodiment, R^(5g) is —CH₂—O—(CH₂)₂—OCH₃, for example R⁵ may be —CH₂—CH—[CH₂—O—(CH₂)₂—OCH₃]-COOH. In another embodiment, R^(5g) is H, for example R⁵ may be —CH₂COOH.

In one embodiment, R⁵ is —C₀₋₃alkylene—C(O)NR^(5h)—CHR^(5l)—COON. The R^(5h) moiety is H or —C₁₋₄alkyl. The R^(5i) moiety is H, —C₁₋₄alkyl, or —C₀₋₃alkylenearyl. In one embodiment, R^(5h) is H and R⁵⁰ is —C₀₋₃alkylenearyl, and the aryl is optionally substituted with 1 to 3 substituents such as —OH, for example, R⁵ may be —C(O)NH—CH(CH₂-phenyl-OH)(COOH).

In another embodiment, R⁵ is —C₀₋₃alkylene-S—SR^(5j), and R^(5j) is selected from —C₁₋₆alkyl, aryl, and —CH₂CH(NH₂)COOH. Examples of such R⁵ groups include —C₀₋₃alkylene-S—S—CH₃, —C₀₋₃alkylene-S—S-phenyl, and —C₀₋₃alkylene-S—S—CH₂CH(NH₂)—COOH.

R⁶ is selected from —C₁₋₆alkyl, —CH₂—O—(CH₂)₂OCH₃, —C₁₋₆alkylene-O—C₁₋₆alkyl, —C₀₋₃alkylenearyl, —C₀₋₃alkyleneheteroaryl, and —C₀₋₃alkylene-C₃₋₇cycloalkyl. In one particular embodiment, R⁶ is selected from —C₁₋₆alkyl, —C₀₋₃alkylenearyl, and —C₀₋₃alkylene-C₃₋₇cycloalkyl. In one particular embodiment, R⁶ is selected from —C₁₋₆alkyl, —C₀₋₃alkylenearyl, and —C₀₋₃alkylene-C₃₋₇cycloalkyl. Each alkyl and each aryl in R⁶ is optionally substituted with 1 to 7 fluoro atoms, where the term “alkyl” is intended to include divalent alkylene groups such as those present in —C₁₋₆alkylene-O—C₁₋₆alkyl and —C₀₋₃alkylene-C₃₋₇cycloalkyl, for example. In addition, each aryl and heteroaryl in R⁶ may be substituted with 1 to 3-OH, —C₁₋₆alkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl, —CN, halo, —O—C₁₋₆alkyl, —S—C₁₋₆alkyl, —S(O)—C₁₋₆alkyl, —S(O)₂—C₁₋₄alkyl, -phenyl, —NO₂, —NH₂, —NH—C₁₋₆alkyl, or —N(C₁₋₆alkyl)₂ groups. Further, each of the aforementioned alkyl, alkenyl and alkynyl groups may be substituted with 1 to 5 fluoro atoms.

In one embodiment, R⁶ is —C₁₋₆alkyl, for example, —CH₃, —CH₂CH₃, —CH(CH₃)₂, —(CH₂)₂CH₃, —(CH₂)₃, —(CH₂)₂CH(CH₃)₂, —CH(CH₃)CH₂CH₃, —CH₂CH(CH₃)₂, —CH₂C(CH₃)₃, and —(CH₂)₄—CH₃. As noted above, each alkyl in R⁶ is optionally substituted with 1 to 7 fluoro atoms. Examples of such fluoro-substituted R⁶ groups include —(CH₂)₂CF₃ and —(CH₂)₃CF₃.

In another embodiment, R⁶ is —CH₂—O—(CH₂)₂OCH₃. In still another one embodiment, R⁶ is —C₁₋₆alkylene-O—C₁₋₆alkyl, for example, —OCH₃ and —CH₂OCH₃.

In one embodiment, R⁶ is —C₀₋₃alkylenearyl, for example, phenyl, benzyl, —CH₂-biphenyl, —(CH₂)₂-phenyl and —CH₂-naphthalen-1-yl. The aryl may be substituted with 1 to 3 substituents. Examples of mono-substituted R⁶ aryl groups include: methylbenzyl such as 4-methylbenzyl; fluoro-substituted alkylbenzyl groups such as 2-trifluoromethylbenzyl; chlorobenzyl such as 2-chlorobenzyl, 3-chlorobenzyl, and 4-chlorobenzyl; fluorobenzyl such as 2-fluorobenzyl, 3-fluorobenzyl, and 4-fluorobenzyl; fluorophenyl; bromobenzyl such as 4-bromobenzyl; iodobenzyl; -benzyl-CF₃, 2-trifluoromethyl-benzyl; -benzyl-CN; and -benzyl-NO₂. Examples of di-substituted R⁶ aryl groups include: di-chlorobenzyl such as 2,4-dichlorobenzyl; and di-fluorobenzyl such as 3,5-difluorobenzyl. Each aryl may also be substituted with 1 to 7 fluoro atoms. Thus, other examples of R⁶ include penta-fluorobenzyl.

In one embodiment, R⁶ is —C₀₋₃alkyleneheteroaryl, for example, —CH₂-Pyridyl, —CH₂-furanyl, —CH₂-thienyl, and —CH₂-thiophenyl such as —CH₂-thiophen-2-yl. In another embodiment, R⁶ is —C₀₋₃alkylene-C₃₋₇cycloalkyl, for example, —CH₂-cyclopropyl, cyclopentyl, —CH₂-cyclopentyl, and —CH₂-cyclohexyl.

R⁷ is H or is taken together with R⁶ to form —C₃₋₈cycloalkyl. In one embodiment, R⁷ is H. In another embodiment, R⁷ is taken together with R⁶ to form —C₃₋₈cycloalkyl, for example cyclopentyl.

One particular embodiment of the invention provides for an active compound of formula I where Ar**—COOH represents Ar—R′ and R⁵ is —C₀₋₃alkylene-SH. One corresponding prodrug (prodrug A) can contain a thioester linkage, which can be cleaved in vivo to form the —COOH(R′) and —C₀₋₃alkylene-SH(R⁵) moieties. Another corresponding prodrug (prodrug B, where Z is —C₁₋₆alkylene, optionally substituted with one or more moieties such as hydroxyl, phenyl, carboxyl, and so forth), contains both an ester and a thioester group, which can be similarly cleaved in vivo, but which also releases a physiologically acceptable acid such as α-hydroxy acid (Z is —CH₂—), β-hydroxy acid (Z is —(CH₂)₂—), (R)-2-hydroxypropionic or lactic acid (Z is —CH(CH₃)—), (R)-hydroxyphenyl acetic or mandelic acid (Z is —CH(phenyl)-), salicylic acid (Z is -phenylene-), 2,3-dihydroxysuccinic or tartaric acid (Z is —CH[CH(OH)(COOH)]-), citric acid (Z is —C[CH₂COOH]_(2—)), hydroxy bis- and hydroxy-tris acids, and so forth.

Yet another corresponding prodrug (prodrug C) is a dimer form of prodrug A, thus containing two thioester linkages, which can both be cleaved in vivo to form two active moieties, each containing the —COOH(R′) and —C₀₋₃alkylene-SH(R⁵) moieties.

Another embodiment of the invention provides for an active compound of formula I where R⁵ is —C₀₋₃alkylene-SH, and the prodrug (prodrug D) is a dimer form of the compound:

Examples of prodrug D include:

For R^(1a)=t-butyl, MS m/z: [M+H⁺] calcd for C₇₈H₈₄N₆O₆S₂, 1,265.59. found 1265.8. For R^(1a)═H, MS m/z: [M+H⁺] calcd for C₇₀H₆₈N₆O₆S₂, 1,153.46. found 1153.6.

In one particular embodiment, the compound of formula I is the species embodied in formula Ia:

where r, n, X, R¹⁻³, and R⁵⁻⁷ are as defined for formula I. In one particular embodiment of the species of formula Ia: r is 1; R¹ is —COOR^(1a), —SO₂NHR^(1d), or tetrazol-5-yl; R^(1a) is H, —C₁₋₆ alkyl, —C₁₋₃ alkylenearyl, —CH(C₁₋₄alkyl)OC(O)R^(1aa), —C₀₋₆ alkylenemorpholine, or

R^(1aa) is —O—C₁₋₆alkyl or —O—C₃₋₇cycloalkyl; R^(1c) is —C₁₋₆alkyl, —C₀₋₆alkylene-O—R^(1ca), —C₁₋₅ alkylene-NR^(1cb)R^(1cc), or —C₀₋₄alkylenearyl; R^(1ca) is H, —C₁₋₆alkyl, or —C₁₋₆alkylene-O—C₁₋₆alkyl; R^(1cb) and R^(1cc) are independently selected from H and —C₁₋₆alkyl, or are taken together as —(CH₂)₂—O—(CF₁₂)₂— or —(CH₂)₂—N[C(O)CH₃]-(CH₂)₂—; R^(1d) is —C(O)R^(1c) or —C(O)NHR^(1c); n is 0 or 1; R² is halo, —C₁₋₆alkyl, or —C₀₋₅alkylene-OR^(2b), where R^(2b) is H; R³ is —C₁₋₁₀alkyl or —C₀₋₅alkylene-O—C₀₋₅alkylene-R^(3b), where R^(ab) is —C₁₋₆alkyl; R⁵ is —C₀₋₃ alkylene-SR^(5a), —C₀₋₃alkylene—C(O)NR^(5b)R^(5c), or —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d); R^(ya) is H or —C(O)—R^(5aa); R^(5aa) is —C₁₋₆alkyl, —C₀₋₆alkylene-C₃₋₇cycloalkyl, -aminoC₄₋₇cycloalkyl, —C₀₋₆alkylenearyl, —C₀₋₆alkyleneheteroaryl, —C₀₋₆alkylenemorpholine, —C₀₋₆alkylenepiperazine—CH₃, —C₀₋₆ alkylenepiperidine, —C₀₋₆alkylenepiperidine—CH₃, —CH[N(R^(5ab))₂]-aa where aa is an amino acid side chain, —C₀₋₆alkylene-CH[N(R^(5ab))₂]—R^(5ac), -2-pyrrolidine, -2-tetrahydrofuran, —C₀₋₆alkylene-OR^(5ab), —C₁₋₂ alkylene-OC(O)—C₁₋₆ alkyl, —C₁₋₄ alkylene-COOH, or -arylene-COOH; R^(5ab) is independently H or —C₁₋₆alkyl; R^(5ac) is H, —C₁₋₆alkyl, —CH₂—C₃₋₇cycloalkyl or —COOH; R^(5b) is —OH; R^(5c) is H; R^(5d) is H or —C₁₋₄alkyl; R⁶ is —C₁₋₆alkyl, —C₀₋₃alkylenearyl, —C₀₋₃alkyleneheteroaryl, or —C₀₋₃alkylene-C₃₋₇cycloalkyl; and R⁷ is H; each alkyl in R² is optionally substituted with 1 to 7 fluoro atoms; each ring in Ar and each aryl and heteroaryl in R¹ and R⁵⁻⁶ is optionally substituted with 1 to 2 substituents independently selected from —C₁₋₆alkyl, halo, —O—C₁₋₆alkyl, and —NH₂, wherein each alkyl is optionally substituted with 1 to 5 fluoro atoms.

When X is selected from —CH₂—NHC(O)—, —NHC(O)—, and —C(O)NH—, the compounds of formula Ia may be depicted as formulas Ia-1, Ia-2, and Ia-3, respectively:

where r, n, R¹⁻³, and R⁵⁻⁶ are as defined for formula I; and pharmaceutically acceptable salts thereof.

In one particular embodiment of the species of formula Ia-1 and Ia-b: r is 1; R¹ is —COOR^(1a) or tetrazol-5-yl, where R^(1a) is H or —C₁₋₆alkyl; n is 1; R² is —C₁₋₆alkyl; R³ is —C₁₋₁₀alkyl; R⁵ is —C₀₋₃alkylene-SR^(5a); R^(5a) is H or —C(O)—R^(5aa), where R^(5aa) is —C₁₋₆alkyl; R⁶ is —C₁₋₆alkyl, —C₀₋₃alkylenearyl, or —C₀₋₃alkylene-C₃₋₇cycloalkyl; and R⁷ is H.

In one particular embodiment of the species of formula Ia-3: r is 1; R¹ is —COOR″, —SO₂NHR^(1d), or tetrazol-5-yl; R^(1a) is H, —C₁₋₆alkyl, —C₁₋₃alkylenearyl, —CH(C₁₋₄alkyl)O—C(O)R^(1aa), —C₀₋₆alkylenemorpholine, or

R^(1aa) is —O—C₁₋₆alkyl or —O—C₃₋₇cycloalkyl; R^(1c) is —C₁₋₆alkyl, —C₀₋₆alkylene-O—R^(1ca), —C₁₋₅alkylene-NR^(1cb)R^(1cc), or —C₀₋₄alkylenearyl; R^(1ca) is H, —C₁₋₆alkyl or —C₁₋₆alkylene-O—C₁₋₆alkyl; R^(aab) and R^(1cc) are independently selected from H and —C₁₋₆alkyl, or are taken together as —(CH₂)₂—O—(CH₂)₂— or —(CH₂)₂—N[C(O)CH₃]-(CH₂)₂—; R^(1d) is —C(O)R^(1c) or —C(O)NHR^(1c); n is 0 or 1; R² is halo, —C₁₋₆alkyl, or —C₀₋₅alkylene-OR^(2b), where Rb is H; R³ is —C₁₋₁₀alkyl or —C₀₋₅alkylene-O—C₀₋₅alkylene-R^(3b), where R^(ab) is —C₁₋₆alkyl; R⁵ is —C₀₋₃alkylene-SR^(5a), —C₀₋₃alkylene—C(O)NR^(5b)R^(5c), or —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d); R^(5a) is H or —C(O)—R^(5aa); R^(5aa) is —C₁₋₆alkyl, —C₀₋₆alkylene-C₃₋₇cycloalkyl, -aminoC₄₋₇cycloalkyl, —C₀₋₆alkylenearyl, —C₀₋₆alkyleneheteroaryl, —C₀₋₆alkylenemorpholine, —C₀₋₆alkylenepiperazine—CH₃, —C₀₋₆alkylenepiperidine, —C₀₋₆alkylenepiperidine—CH₃, —CH[N(R^(5ab))₂]-aa where aa is an amino acid side chain, —C₀₋₆alkylene-CH[N(R^(5ab))₂]—R^(5ac), -2-pyrrolidine, -2-tetrahydrofuran, —C₀₋₆alkylene-OR^(5ab), —C₁₋₂alkylene-OC(O)—C₁₋₆alkyl, —C₁₋₄alkylene-COOH, or -arylene-COOH; R^(5ab) is independently H or —C₁₋₆alkyl; R^(5aa) is H, —C₁₋₆alkyl, —CH₂—C₃₋₇cycloalkyl or —COOH; R^(5b) is —OH; R^(5a) is H; R^(5d) is H or —C₁₋₄alkyl; R⁶ is —C₁₋₆alkyl, —C₀₋₃alkylenearyl, —C₀₋₃alkyleneheteroaryl, or —C₀₋₃alkylene-C₃₋₇cycloalkyl; and R⁷ is H; each alkyl in R² is optionally substituted with 1 to 7 fluoro atoms; each ring in Ar and each aryl and heteroaryl in R¹ and R⁵⁻⁶ is optionally substituted with 1 to 2 substituents independently selected from —C₁₋₆alkyl, halo, —O—C₁₋₆alkyl, and —NH₂, wherein each alkyl is optionally substituted with 1 to 5 fluoro atoms.

In another embodiment of the species of formula Ia-3: r is 1; R¹ is —SO₂NHR^(1d); R^(1c) is —C₁₋₆alkyl, —C₀₋₆alkylene-O—R^(1ca), —C₁₋₅alkylene-NR^(1cb)R^(1cc), or —C₀₋₄alkylenearyl; R^(1ca) is H, —C₁₋₆alkyl, or —C₁₋₆alkylene-O—C₁₋₆alkyl; R^(1cb) and R^(1cc) are independently selected from H and —C₁₋₆alkyl, or are taken together as —(CH₂)₂—O—(CH₂)₂— or —(CH₂)₂—N[C(O)CH₃]—(CH₂)₂—; R^(1d) is —C(O)R^(1c) or —C(O)NHR^(1c); n is 1; R² is —C₁₋₆alkyl; R³ is —C₁₋₁₀alkyl; R⁵ is —C₀₋₃alkylene-SR^(5a) or —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d); R^(5a) is H or —C(O)—R^(5aa); R^(5aa) is —C₁₋₆alkyl; R^(5b) is —OH; R^(5d) is H; R⁶ is —C₁₋₆alkyl or —C₀₋₃alkylenearyl; and R⁷ is H.

In one particular embodiment, the compound of formula I is the species embodied in formula Ib:

In one particular embodiment of the species of formula Ib: r is 1; R¹ is —COOR^(1a), where R^(1a) is H or —C₁₋₆alkyl; n is 1; R² is —C₁₋₆alkyl; R³ is —C₁₋₁₀alkyl; X is —CH₂—NHC(O)—, —NHC(O)—, or —C(O)NH—; R⁵ is —C₀₋₃alkylene-SR^(5a), —C₀₋₃alkylene—C(O)NR^(5b)R^(5c), or —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d); R^(5a) is H or —C(O)—R^(5aa); R^(5aa) is —C₁₋₆alkyl; R^(5b) is —OH; R^(5a) is H; R^(5d) is H or —C₁₋₄alkyl; R⁶ is —C₁₋₆alkyl, —C₀₋₃alkylenearyl, or —C₀₋₃alkylene-C₃₋₇cycloalkyl; R⁷ is H; the alkyl in R⁶ is optionally substituted with 1 to 7 fluoro atoms; and the phenyl ring is optionally substituted with 1 halo atom.

When X is selected from —CH₂—NHC(O)—, —NHC(O)—, and —C(O)NH—, the compounds of formula Ib may be depicted as formulas Ib-1, Ib-2, and Ib-3, respectively:

where r, n, R¹⁻³, and R⁵⁻⁶ are as defined for formula I; and pharmaceutically acceptable salts thereof.

In one particular embodiment of the species of formula Ib-1: r is 1; R¹ is —COOR^(1a), where R^(1a) is H or —C₁₋₆alkyl; n is 1; R² is —C₁₋₆alkyl; R³ is —C₁₋₁₀alkyl; R⁵ is —C₀₋₃alkylene-SR^(5a) or —C₀₋₃alkylene—C(O)NR^(5b)R^(5c); R^(5a) is H or —C(O)—R^(5 aa); R^(5aa) is —C₁₋₆alkyl; R^(5b) is —OH; R^(5a) is H; R⁶ is —C₁₋₆alkyl, —C₀₋₃alkylenearyl, or —C₀₋₃alkylene-C₃₋₇cycloalkyl; R⁷ is H; the alkyl in R⁶ is optionally substituted with 1 to 7 fluoro atoms; and the phenyl ring is optionally substituted with 1 halo atom.

In one particular embodiment of the species of formula Ib-2: r is 1; R¹ is —COOR^(1a), where R^(1a) is H or —C₁₋₆alkyl; n is 1; R² is —C₁₋₆alkyl; R³ is —C₁₋₁₀alkyl; R⁵ is —C₀₋₃alkylene-SR^(5a), —C₀₋₃alkylene—C(O)NR^(5b)R^(5c), or —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d); R^(5a) is H or —C(O)—R^(5aa); R^(5aa) is —C₁₋₆alkyl; R^(5b) is —OH; R^(5a) is H; R^(5d) is H or —C₁₋₄alkyl; R⁶ is —C₁₋₆alkyl, —C₀₋₃alkylenearyl, or —C₀₋₃alkylene-C₃₋₇cycloalkyl; R⁷ is H; the alkyl in R⁶ is optionally substituted with 1 to 7 fluoro atoms; and the phenyl ring is optionally substituted with 1 halo atom.

In one particular embodiment of the species of formula Ib-3: r is 1; R¹ is —COOR^(1a), where R^(1a) is H or —C₁₋₆alkyl; n is 1; R² is —C₁₋₆alkyl; R³ is —C₁₋₁₀alkyl; R⁵ is —C₀₋₃alkylene-SR^(5a), —C₀₋₃alkylene—C(O)NR^(5b)R^(5c), or —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d); R^(5a) is H or —C(O)—R^(5aa); R^(5aa) is —C₁₋₆alkyl; R^(5b) is —OH; R^(5a) is H; R^(5d) is H or —C₁₋₄alkyl; R⁶ is —C₁₋₆alkyl or —O₀₋₃alkylenearyl; and R⁷ is H.

In one particular embodiment, the compound of formula I is the species embodied in formula Ic:

In one particular embodiment of the species of formula Ic: r is 1; R¹ is —COOR^(1a) or —SO₂NHR^(1d); R^(1a) is R^(1a) H or —C₁₋₆alkyl; R^(1d) is —C(O)R^(1c); R^(1c) is —C₁₋₆alkyl; n is 1; R² is —C₁₋₆alkyl; R³ is —C₁₋₁₀alkyl; X is —C(O)NH—; R⁵ is —C₀₋₃alkylene-SR^(5a); R^(5a) is H or —C(O)—R^(5aa); R^(5aa) is —C₁₋₆alkyl; R⁶ is —C₁₋₆alkyl or —C₀₋₃alkylenearyl; and R⁷ is H.

In one particular embodiment, the compound of formula I is the species embodied in formula Id:

In one particular embodiment of the species of formula Id: r is 1; R¹ is —COOR^(1a), where R^(1a) is H or —C₁₋₆alkyl; n is 1; R² is —C₁₋₆alkyl; R³ is —C₁₋₁₀alkyl; X is —C(O)NH—; R⁵ is —C₀₋₃alkylene-SR^(5a); R^(5a) is H or —C(O)—R^(5aa); R^(5aa) is —C₁₋₆alkyl; R⁶ is —C₁₋₆alkyl or —C₀₋₃alkylenearyl; and R⁷ is H.

In one particular embodiment, r is 1; and Ar is an aryl group selected from:

where Ar is optionally substituted with one fluoro or chloro atom.

In one particular embodiment, r is 1; and n is 0, or n is 1 and R² is at the 4 position.

In another aspect, these embodiments have formula Ia, Ia-1, Ia-2, Ia-3, Ib, Ib-1, Ib-2, Ib-3, Ic or Id.

In another particular embodiment, R¹ is selected from —COOH, —NHSO₂R^(1b), —SO₂NHR^(1d), —SO₂OH, —C(O)NH—SO₂R^(1c), —P(O)(OH)₂, —CN, —O—CH(R^(1e))—COOH, tetrazol-5-yl,

where R^(1b), R^(1c), R^(1d) and R^(1e), are as defined for formula I. In one particular embodiment, , R¹ is selected from —COOR^(1a), —SO₂NHR^(1d), and tetrazol-5-yl. In another embodiment, R¹ is selected from —COOH, —SO₂NHR^(1d) (for example —SO₂NHC(O)—C₁₋₆alkyl), and tetrazol-5-yl. In another aspect, these embodiments have formula Ia, Ia-1, Ia-2, Ia-3, Ib, Ib-1, Ib-2, Ib-3, Ic or Id.

In one particular embodiment, R¹ is —COOR^(1a), where R^(1a) is —C₁₋₆alkyl, —C₁₋₃alkylenearyl, —C₁₋₃alkyleneheteroaryl, —C₃₋₇cycloalkyl, —CH(C₁₋₄alkyl)OC(O)R^(1aa), —C₀₋₆alkylenemorpholine,

where R^(1aa) is as defined for formula I. In one aspect of the invention, these compounds may find particular utility as prodrugs or as intermediates in the synthetic procedures described herein. In one particular embodiment, R¹ is —COOR^(1a) and R^(1a) is —C₁₋₆alkyl. In another aspect, these embodiments have formula Ia, Ia-1, Ia-2, Ia-3, Ib, Ib-1, Ib-2, Ib-3, Ic or Id.

In one particular embodiment, R¹ is selected from —COOR^(1a) and tetrazol-5-yl, where R^(1a) is H or —C₁₋₆alkyl. In another aspect, this embodiment has formula Ia, Ia-1, Ia-2, Ia-3, Ib, Ib-1, Ib-2, Ib-3, Ic or Id.

In one embodiment, R⁵ is selected from —C₀₋₃alkylene-SR^(5a), —C₀₋₃alkylene-C(O)NR^(5b)R^(5c), —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d), —NH—C₀₋₁alkylene-P(O)(OR^(5e))₂, —C₀₋₃alkylene-P(O)OR^(5e)R^(5f), —C₀₋₂alkylene-CHR^(5g)—COOH, and —C₀₋₃alkylene—C(O)NR^(5h)—CHR^(5l)—COOH; where R^(5a) is H, R^(5b) is —OH, R^(5e) is H, R^(5d) is H, R^(ye) is H; and R^(5f), R^(5g), R^(5aa), R⁵¹ are as defined for formula I. More particularly, in one embodiment, R⁵ is selected from —C₀₋₁alkylene-SH, —C₀₋₁alkylene—C(O)—N(OH)H, and —C₀₋₃alkylene-N(OH)—C(O)H. In another embodiment, R⁵ is selected from —C₀₋₃alkylene-SR^(5a) and —C₀₋₃alkylene-C(O)NR^(5b)R^(5c), where R^(5a) is H and R^(5b) is —OH. In one particular embodiment, R^(5a) is H. In another aspect, these embodiments have formula Ia, Ia-1, Ia-2, Ia-3, Ib, Ib-1, Ib-2, Ib-3, Ic or Id.

In yet another embodiment, R⁵ is selected from —C₀₋₃alkylene-SR^(5a), —C₀₋₃alkylene-C(O)NR^(5b)R^(5c), —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d), —NH—C₀₋₁alkylene-P(O)(OR^(5e))₂, —C₀₋₃alkylene-P(O)OR^(5e)R^(5f), and —C₀₋₃alkylene-S—SR^(5j); where R^(5a) is —C(O)—R^(5aa); R^(5b) is H, —OC(O)R^(5ba), —CH₂COOH, —O-benzyl, -pyridyl, or —OC(S)NR^(5bb)R^(5bc); R^(5e) is —C₁₋₆alkyl, —C₁₋₃alkylenearyl, —C₁₋₃ alkylenehetero aryl, —C₃₋₇ cyclo alkyl, —CH(CH₃)—O—C(O)R^(5ea),

and where R^(5aa), R^(5ba), R^(5bb), R^(5bc), R^(5c), R^(5d), R^(5ea), R^(5f) and R^(5j) are as defined for formula I. In one aspect of the invention, these compounds may find particular utility as prodrugs or as intermediates in the synthetic procedures described herein. In another aspect, these embodiments have formula Ia, Ia-1, Ia-2, Ia-3, Ib, Ib-1, Ib-2, Ib-3, Ic or Id.

In one particular embodiment, R⁵ is selected from —C₀₋₃alkylene-SR^(5a) and —C₀₋₃alkylene—C(O)NR^(5b)R^(5c); where R^(5a) is selected from H and —C(O)—C₁₋₆alkyl; R^(5b) is selected from H, —OH, and —OC(O)—C₁₋₆alkyl; and R^(5a) is selected from H and —C₁₋₆alkyl. In another aspect, this embodiment has formula Ia, Ia-1, Ia-2, Ia-3, Ib, Ib-1, Ib-2, Ib-3, Ic or Id.

In another embodiment, R¹ is selected from —COOR^(1a) where R^(1a) is H, —NHSO₂R^(1b), —SO₂NHR^(1d), —SO₂OH, —C(O)NH—SO₂R^(1c), —P(O)(OH)₂, —CN, —O—CH(R^(1e))—COOH, tetrazol-5-yl,

R⁵ is selected from —C₀₋₃alkylene-SR^(5a), —C₀₋₃alkylene—C(O)NR^(5b)R^(5c), —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d), —NH—C₀₋₁alkylene-P(O)(OR^(5e))₂, —C₀₋₃alkylene-P(O)OR^(5e)R^(5f), —C₀₋₂alkylene-CHR^(5g)—COOH, and —C₀₋₃alkylene—C(O)NR^(5l)—CHR^(5l)—COOH; R^(5a) is H; R^(5b) is —OH; R^(5a) is H; R^(5d) is H; R^(5e) is H; and R^(1b), R^(1c), R^(1d), R^(1e), R^(5f), R^(5g), R^(5h), R^(5i) are as defined for formula I. In one particular embodiment, R¹ is selected from —COOH, —SO₂NHR^(1d), and tetrazol-5-yl; and R⁵ is selected from —C₀₋₃alkylene-SH, and —C₀₋₃alkylene—C(O)N(OH)H. In another aspect, these embodiments have formula Ia, Ia-1, Ia-2, Ia-3, Ib, Ib-1, Ib-2, Ib-3, Ic or Id.

In another embodiment, R¹ is —COOR^(1a), where R^(1a) is selected from —C₁₋₆alkyl, —C₁₋₃alkylenearyl, —C₁₋₃alkyleneheteroaryl, —C₃₋₇cycloalkyl, —CH(C₁₋₄alkyl)OC(O)R^(1aa), —C₀₋₆alkylenemorpholine,

R⁵ is selected from —C₀₋₃alkylene-SR^(5a), —C₀₋₃alkylene—C(O)NR^(5b)R^(5c), —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d), —NH—C₀₋₁alkylene-P(O)(OR^(5e))₂, —C₀₋₃alkylene-P(O)OR^(5e)R^(5f), and —C₀₋₃alkylene-S—SR^(5j); where R^(5a) is —C(O)—R^(5aa); R^(5b) is H, —OC(O)R^(5ba), —CH₂COOH, —O-benzyl, -pyridyl, or —OC(S)NR^(5bb)R^(5bc); R^(5e) is —C₁₋₆alkyl, —C₁₋₃alkylenearyl, —C₁₋₃alkyleneheteroaryl, —C₃₋₇cycloalkyl, —CH(CH₃)—O—C(O)R^(5ea),

where R^(1aa), R^(5aa), R^(5ba), R^(5bb), R^(5bc), R^(5c), R^(5d), R^(5ea), R^(f), and R^(5j) are as defined for formula I. In another aspect, this embodiment has formula Ia, Ia-1, Ia-2, Ia-3, Ib, Ib-1, Ib-2, Ib-3, Ic or Id.

In another particular embodiment, R¹ is selected from —COOR^(1a) where R^(1a) is H, —NHSO₂R^(1b), —SO₂NHR^(1d), —SO₂OH, —C(O)NH—SO₂R^(1c), —P(O)(OH)₂, —CN, —O—CH(R^(1e))—COOH, tetrazol-5-yl,

R⁵ is selected from —C₀₋₃alkylene-SR^(5a), —C₀₋₃alkylene—C(O)NR^(5b)R^(5c), —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d), —NH—C₀₋₁alkylene-P(O)(OR^(5e))₂, —C₀₋₃alkylene-P(O)OR^(5e)R^(5f), and —O₀₋₃alkylene-S—SR^(5a); where R^(5a) is —C(O)—R^(5aa); R^(5b) is H, —OC(O)R^(5ba), —CH₂COOH, —O-benzyl, -pyridyl, or —OC(S)NR^(5bb)R^(5bc); R^(5e) is —C₁₋₆alkyl, —C₁₋₃alkylenearyl, —C₁₋₃alkyleneheteroaryl, —C₃₋₇cycloalkyl, —CH(CH₃)—O—C(O)R^(5ea),

and where R^(1b), R^(1c), R^(1d), R^(1e), R^(5aa), R^(5ba), R^(5bb), R^(5bc), R^(5c), R^(5d), R^(5ea), R^(5f), and R^(5j) are as defined for formula I. In another aspect, this embodiment has formula Ia, Ia-1, Ia-2, Ia-3, Ib, Ib-1, Ib-2, Ib-3, Ic or Id.

In another embodiment, R¹ is —COOR^(1a), where R^(1a) is selected from —C₁₋₆alkyl, —C₁₋₃ alkylenearyl, —C₁₋₃alkyleneheteroaryl, —C₃ ₋₇ cycloalkyl, —CH(C₁₋₄ alkyl)OC(O)R^(1aa), —C₀₋₆alkylenemorpholine,

R⁵ is selected from —C₀₋₃alkylene-SR^(5a), —C₀₋₃alkylene—C(O)NR^(5b)R^(5c), —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d), —NH—C₀₋₁alkylene-P(O)(OR^(5e))₂, —C₀₋₃alkylene-P(O)OR^(5e)R^(5f), —C₀₋₂alkylene-CHR^(5g)—COOH, and —C₀₋₃alkylene—C(O)NR^(5h)—CHR^(5l)—COOH; R^(5a) is H; R^(5b) is —OH; R⁵ is H; R^(5d) is H; R^(5e) is H; and R^(1aa), R^(5f), R^(5g), R^(5h), R^(5i) are as defined for formula I. In another aspect, this embodiment has formula Ia, Ia-1, Ia-2, Ia-3, Ib, Ib-1, Ib-2, Ib-3, Ic or Id.

A particular group of compounds of formula I are those disclosed in U.S. Provisional Application No. 60/933,207, filed on Jun. 5, 2007. This group includes compounds of formula I′:

wherein: r′ is 0, 1 or 2; Ar′ is an aryl group selected from:

R¹′ is selected from —COOR^(1a)′ —NHSO₂—C₁₋₆alkyl, —NHSO₂aryl, —NHSO₂NHC(O)—C₁₋₆alkyl, —NHSO₂NHC(O)-aryl, —SO₂NHC(O)—C₁₋₆alkyl, —SO₂NHC(O)-aryl, —SO₂NHC(O)NH—C₁₋₆alkyl, —SO₂NHC(O)NH-aryl, —SO₂OH, —SO₂NH₂, —SO₂NH—C₁₋₆alkyl, —SO₂NH-aryl, —C(O)NH—SO₂—C₁₋₆alkyl, —C(O)NH—SO₂-aryl, —P(O)(OH)₂, —CN, —OCH(CH₃)—COOH, —OCH(aryl)-COOH, tetrazol-5-yl,

where R^(1a)′ is selected from H, —C₁₋₆alkyl, benzyl, —C₁₋₃alkyleneheteroaryl, cycloalkyl, —CH(CH₃)OC(O)R^(1b)′,

R^(1b)′ is selected from —O—C₁₋₆alkyl, —O-cycloalkyl, —NR^(1c)′R^(1d)′, —CH(NH₂)CH₂COOCH₃; and R^(1c)′ and R^(1d)′ are independently selected from H, —C₁₋₆alkyl, and benzyl, or are taken together as —(CH₂)₃₋₆—;

n′ is 0, 1, 2 or 3; R²′ is selected from —CH₂OH, halo, —NO₂, —C₁₋₆alkyl, —C₂₋₆alkenyl, —C₃₋₆cycloalkyl, —CN, —C(O)R²″, —C₀₋₅alkylene-OR^(2b)v, —C₀₋₅alkylene-NR^(2c)′R^(2d)′, —C₀₋₃alkylenearyl, and —C₀₋₃alkyleneheteroaryl; where R^(2a)′ is selected from H, —C₁₋₆alkyl, —C₃₋₆cycloalkyl, —C₀₋₃alkylenephenyl, —OR^(2b)′, and —NR^(2c)′R^(2d)′; R^(2b)′ is selected from H, —C₁₋₆alkyl, —C₃₋₆cycloalkyl, and —C₀₋₁alkylenephenyl; and R^(2c)′ and R^(2d)′ are independently selected from H, —C₁₋₄alkyl, and —C₀₋₁alkylenephenyl;

R³′ is selected from —C₁₋₁₀alkyl, —C₂₋₁₀alkenyl, —C₃₋₁₀alkynyl, —C₀₋₃alkylene-C₃₋₇cycloalkyl, —C₂₋₃alkenylene-C₃₋₇cycloalkyl, —C₂₋₃alkynylene-C₃₋₇cycloalkyl, —C₀₋₅alkylene-NR³′—C₀₋₅alkylene-R^(3b)′, —C₀₋₅alkylene-O—C₁₋₅alkylene-R^(3b)′, —C₁₋₅alkylene-S—C₁₋₅alkylene-R^(3b)′ and —C₀₋₃alkylenearyl; where R^(3a)′ is selected from H, —C₁₋₆alkyl, —C₃₋₆cycloalkyl, and —C₀₋₁alkylenephenyl; and R^(3b)′ is selected from H, —C₁₋₆alkyl, —C₃₋₆cycloalkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl, and phenyl;

X′ is —C₁₋₁₂alkylene-, where at least one —CH₂— moiety in the alkylene is replaced with a —NR^(4a)′—C(O)— or —C(O)—NR⁴′-moiety, where R^(4a)′ is selected from H, —OH, and —C₁₋₄alkyl;

R⁵′ is selected from —C₀₋₃ alkylene-SR^(5a)′, —C₀₋₃alkylene—C(O)NR^(5b)′R^(5c)′, —C₀₋₃alkylene-NR^(5b)′—C(O)R^(5d)′, —C₀₋₁alkylene-NHC(O)CH₂SH, —NH—C₀₋₁alkylene-P(O)(OR^(5e)′)₂, —C₀₋₃alkylene-P(O)OR^(5e)′R^(5f)′, —C₀₋₂alkylene-CHR^(5g)′-COOH and —C₀₋₃alkylene—C(O)NR^(5h)′—CHR^(5i)′—COON; where R^(5a)′ is selected from H, —C(O)—C₁₋₆alkyl, —C(O)—C₀₋₆alkylene-C₃₋₇cycloalkyl, —C(O)—C₀₋₆alkylenearyl, —C(O)—C₀₋₆alkyleneheteroaryl, —C(O)—C₁₋₆alkyl, —C(O)—OC₀₋₆alkylenearyl, —C₁₋₂alkylene-OC(O)—C₁₋₆alkyl, —C₁₋₂alkylene-OC(O)—C₀₋₆alkylenearyl, and —C₁₋₂alkylene-OC(O)—OC₁₋₆alkyl; R^(5b)′ is selected from H, —OH, —OC(O)—C₁₋₆alkyl, —CH₂COOH, —O-benzyl, -pyridyl, —OC(O)OCH₂-phenyl, —OC(O)CH₂O-phenyl, —OC(O)N(CH₃)₂, and —OC(S)N(CH₃)₂; R^(5c)′ is selected from H, —C₁₋₆alkyl, and —C(O)—R^(5c)″, where R^(5c)″ is selected from —C₁₋₆alkyl, —C₃₋₇cycloalkyl, aryl, and heteroaryl; R^(5d)′ is selected from H, —C₀₋₃alkylenearyl, —NR^(5d)″R^(5d)′″, and —O—C₁₋₆alkyl, where R^(5d)″ and R^(5d)′″ are independently selected from H and —C₁₋₄alkyl; R^(5e)′ is selected from H, —C₁₋₆alkyl, benzyl, —C₁₋₃alkyleneheteroaryl, cycloalkyl, —CH(CH₃)OC(O)R^(5e)″,

where R^(5e)″ is selected from —O—C₁₋₆alkyl, —O-cycloalkyl, —NR^(5e)′″R^(5e)′″, and —CH(NH₂)CH₂COOCH₃, and where R^(5e)′″ and R^(5e)″″ are independently selected from H, —C₁₋₆alkyl, and benzyl, or are taken together as —(CH₂)₃₋₆—; R^(5f′) is selected from H, —C₁₋₄ alkyl, —C₀₋₃alkylenearyl, —C₁₋₃alkylene-NR^(5f)″R^(5aa), and —C₁₋₃alkylene(aryl)-C₀₋₃alkylene-NR^(5f)″R^(5aa), where R^(5f)″ and R^(f)′″ are independently selected from H and —C₁₋₄alkyl; R^(59g)′ is selected from H, —C₁₋₆alkyl, —C₁₋₃alkylenearyl, and —CH₂—O—(CH₂)₂—OCH₃; R^(5h)′ is selected from H and —C₁₋₄alkyl; and R^(5l)′ is selected from H, —C₁₋₄alkyl, and —C₀₋₃alkylenearyl;

R⁶′ is selected from —C₁₋₆alkyl, —CH₂—O—(CH₂)₂OCH₃, —C₁₋₆alkylene-O—C₁₋₆alkyl, —C₀₋₃alkylenearyl, —C₀₋₃alkyleneheteroaryl, and —C₀₋₃alkylene-C₃₋₇cycloalkyl; and

R⁷′ is H or is taken together with R⁶′ to form —C₃₋₈cycloalkyl; wherein: each —CH₂— group in —(CH₂)_(r)— is optionally substituted with 1 or 2 substituents independently selected from —C₁₋₄alkyl and fluoro; each carbon atom in the alkylene moiety in X′ is optionally substituted with one or more R^(4b)′ groups and one —CH₂— moiety in X′ may be replaced with —C₄₋₈cycloalkylene-; wherein R^(4b)′ is selected from —C₀₋₅alkylene-COOH, —C₁₋₆alkyl, —C₀₋₁alkylene-CONH₂, —C₁₋₂alkylene-OH, —C₀₋₃alkylene-C₃₋₇cycloalkyl, and benzyl; each alkyl and each aryl in R¹′⁻³′, R^(4a)′^(-4b)′, and R⁵′⁻⁶′ is optionally substituted with 1 to 7 fluoro atoms; each ring in Ar′ and each aryl in R¹′⁻³′ and R⁵′⁻⁶′ is optionally substituted with 1 to 3 substituents independently selected from —OH, —C₁₋₆alkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl, —CN, halo, —O—C₁₋₆alkyl, —S—C₁₋₆alkyl, —S(O)—C₁₋₆alkyl, —S(O)₂—C₁₋₄alkyl, -phenyl, —NO₂, —NH₂, —NH—C₁₋₆alkyl and —N(C₁₋₆alkyl)₂, wherein each alkyl, alkenyl and alkynyl is optionally substituted with 1 to 5 fluoro atoms; and pharmaceutically acceptable salts thereof.

In addition, particular compounds of formula I that are of interest include those set forth in the Examples below, as well as the pharmaceutically acceptable salts thereof.

DEFINITIONS

When describing the compounds, compositions, methods and processes of the invention, the following terms have the following meanings unless otherwise indicated.

Additionally, as used herein, the singular forms “a,” “an,” and “the” include the corresponding plural forms unless the context of use clearly dictates otherwise. The terms “comprising”, “including,” and “having” are intended to be inclusive and mean that there may be additional elements other than the listed elements.

The term “pharmaceutically acceptable” refers to a material that is not biologically or otherwise undesirable. For example, the term “pharmaceutically acceptable carrier” refers to a material that can be incorporated into a composition and administered to a patient without causing undesirable biological effects or interacting in a deleterious manner with other components of the composition. Such pharmaceutically acceptable materials typically have met the required standards of toxicological and manufacturing testing, and include those materials identified as suitable inactive ingredients by the U.S. Food and Drug Administration.

The term “pharmaceutically acceptable salt” means a salt prepared from a base or an acid which is acceptable for administration to a patient, such as a mammal (for example, salts having acceptable mammalian safety for a given dosage regime). However, it is understood that the salts covered by the invention are not required to be pharmaceutically acceptable salts, such as salts of intermediate compounds that are not intended for administration to a patient. Pharmaceutically acceptable salts can be derived from pharmaceutically acceptable inorganic or organic bases and from pharmaceutically acceptable inorganic or organic acids. In addition, when a compound of formula I contains both a basic moiety, such as an amine, pyridine or imidazole, and an acidic moiety such as a carboxylic acid or tetrazole, zwitterions may be formed and are included within the term “salt” as used herein. Salts derived from pharmaceutically acceptable inorganic bases include ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic, manganous, potassium, sodium, and zinc salts, and the like. Salts derived from pharmaceutically acceptable organic bases include salts of primary, secondary and tertiary amines, including substituted amines, cyclic amines, naturally-occurring amines and the like, such as arginine, betaine, caffeine, choline, N,N′-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine and the like. Salts derived from pharmaceutically acceptable inorganic acids include salts of boric, carbonic, hydrohalic (hydrobromic, hydrochloric, hydrofluoric or hydroiodic), nitric, phosphoric, sulfamic and sulfuric acids. Salts derived from pharmaceutically acceptable organic acids include salts of aliphatic hydroxyl acids (for example, citric, gluconic, glycolic, lactic, lactobionic, malic, and tartaric acids), aliphatic monocarboxylic acids (for example, acetic, butyric, formic, propionic and trifluoroacetic acids), amino acids (for example, aspartic and glutamic acids), aromatic carboxylic acids (for example, benzoic, p-chlorobenzoic, diphenylacetic, gentisic, hippuric, and triphenylacetic acids), aromatic hydroxyl acids (for example, o-hydroxybenzoic, p-hydroxybenzoic, 1-hydroxynaphthalene-2-carboxylic and 3-hydroxynaphthalene-2-carboxylic acids), ascorbic, dicarboxylic acids (for example, fumaric, maleic, oxalic and succinic acids), glucoronic, mandelic, mucic, nicotinic, orotic, pamoic, pantothenic, sulfonic acids (for example, benzenesulfonic, camphosulfonic, edisylic, ethanesulfonic, isethionic, methanesulfonic, naphthalenesulfonic, naphthalene-1,5-disulfonic, naphthalene-2,6-disulfonic and p-toluenesulfonic acids), xinafoic acid, and the like.

As used herein, the term “prodrug” is intended to mean an inactive (or significantly less active) precursor of a drug that is converted into its active form in the body under physiological conditions, for example, by normal metabolic processes. The term is also intended to include certain protected derivatives of compounds of formula I that may be made prior to a final deprotection stage. Such compounds may not possess pharmacological activity at AT₁ and/or NEP, but may be administered orally or parenterally and thereafter metabolized in the body to form compounds of the invention which are pharmacologically active at AT₁ and/or NEP. Thus, all protected derivatives and prodrugs of compounds formula I are included within the scope of the invention. Prodrugs of compounds of formula I having a free carboxyl, sulfhydryl or hydroxy group can be readily synthesized by techniques that are well known in the art. These prodrug derivatives are then converted by solvolysis or under physiological conditions to be the free carboxyl, sulfhydryl and/or hydroxy compounds. Exemplary prodrugs include: esters including C₁₋₆alkylesters and aryl-C₁₋₆alkylesters, carbonate esters, hemi-esters, phosphate esters, nitro esters, sulfate esters, sulfoxides, amides, carbamates, azo-compounds, phosphamides, glycosides, ethers, acetals, ketals, and disulfides. In one embodiment, the compounds of formula I have a free sulfhydryl or a free carboxyl and the prodrug is an ester derivative thereof, i.e., the prodrug is a thioester such as —SC(O)CH₃ or an ester such as —C(O)OCH₃.

The term “solvate” means a complex or aggregate formed by one or more molecules of a solute, for example, a compound of formula I or a pharmaceutically acceptable salt thereof, and one or more molecules of a solvent. Such solvates are typically crystalline solids having a substantially fixed molar ratio of solute and solvent. Representative solvents include, by way of example, water, methanol, ethanol, isopropanol, acetic acid and the like. When the solvent is water, the solvate formed is a hydrate.

The term “therapeutically effective amount” means an amount sufficient to effect treatment when administered to a patient in need thereof, that is, the amount of drug needed to obtain the desired therapeutic effect. For example, a therapeutically effective amount for treating hypertension could be the amount of drug needed to reduce blood pressure or the amount of drug needed to maintain normal blood pressure. On the other hand, the term “effective amount” means an amount sufficient to obtain a desired result, which may not necessary be a therapeutic result. For example, when studying a system comprising an AT₁ receptor, an “effective amount” may be the amount needed to antagonize the receptor.

The term “treating” or “treatment” as used herein means the treating or treatment of a disease or medical condition (such as hypertension) in a patient, such as a mammal (particularly a human) that includes: (a) preventing the disease or medical condition from occurring, that is, prophylactic treatment of a patient; (b) ameliorating the disease or medical condition such as by eliminating or causing regression of the disease or medical condition in a patient; (c) suppressing the disease or medical condition such as by slowing or arresting the development of the disease or medical condition in a patient; or (d) alleviating the symptoms of the disease or medical condition in a patient. For example, the term “treating hypertension” would include preventing hypertension from occurring, ameliorating hypertension, suppressing hypertension, and alleviating the symptoms of hypertension (for example, lowering blood pressure). The term “patient” is intended to include those mammals, such as humans, that are in need of treatment or disease prevention, or that are presently being treated for disease prevention or treatment of a specific disease or medical condition. The term “patient” also includes test subjects in which compounds of the invention are being evaluated or test subjects are being used in a assay, for example an animal model.

The term “alkyl” means a monovalent saturated hydrocarbon group which may be linear or branched. Unless otherwise defined, such alkyl groups typically contain from 1 to 10 carbon atoms and include, for example, —C₁₋₄alkyl, —C₁₋₆alkyl, and —C₁₋₁₀alkyl. Representative alkyl groups include, by way of example, methyl, ethyl, n-propyl, isopropyl, n-butyl, s-butyl, isobutyl, t-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl and the like.

When a specific number of carbon atoms is intended for a particular term used herein, the number of carbon atoms is shown preceding the term as subscript. For example, the term “—C₁₋₆alkyl” means an alkyl group having from 1 to 6 carbon atoms, and the term “—C₃₋₆cycloalkyl” means a cycloalkyl group having from 3 to 6 carbon atoms, where the carbon atoms are in any acceptable configuration.

The term “alkylene” means a divalent saturated hydrocarbon group that may be linear or branched. Unless otherwise defined, such alkylene groups typically contain from 0 to 12 carbon atoms and include, for example, —C₀₋₁alkylene-, —C₀₋₂alkylene-, —C₀₋₃alkylene-, —C₀₋₅alkylene-, —C₀₋₆alkylene-, —C₁₋₂alkylene- and —C₁₋₁₂alkylene-. Representative alkylene groups include, by way of example, methylene, ethane-1,2-diyl (“ethylene”), propane-1,2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl and the like. It is understood that when the alkylene term include zero carbons such as —C₀₋₁alkylene- or —C₀₋₅alkylene-, such terms are intended to include a single bond.

The term “alkylthio” means a monovalent group of the formula —S-alkyl, where alkyl is as defined herein. Unless otherwise defined, such alkylthio groups typically contain from 1 to 10 carbon atoms and include, for example, —S—C₁₋₄alkyl and —S—C₁₋₆alkyl. Representative alkylthio groups include, by way of example, ethylthio, propylthio, isopropylthio, butylthio, s-butylthio and t-butylthio.

The term “alkenyl” means a monovalent unsaturated hydrocarbon group which may be linear or branched and which has at least one, and typically 1, 2 or 3, carbon-carbon double bonds. Unless otherwise defined, such alkenyl groups typically contain from 2 to 10 carbon atoms and include, for example, —C₂₋₄alkenyl and —C₂₋₁₀alkenyl. Representative alkenyl groups include, by way of example, ethenyl, n-propenyl, isopropenyl, n-but-2-enyl, n-hex-3-enyl and the like. The term “alkenylene” means a divalent alkenyl group, and includes groups such as —C₂₋₃alkenylene-.

The term “alkoxy” means a monovalent group of the formula —O-alkyl, where alkyl is as defined herein. Unless otherwise defined, such alkoxy groups typically contain from 1 to 10 carbon atoms and include, for example, —O—C₁₋₄alkyl and —O—C₁₋₆alkyl. Representative alkoxy groups include, by way of example, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, isobutoxy, t-butoxy and the like.

The term “alkynyl” means a monovalent unsaturated hydrocarbon group which may be linear or branched and which has at least one, and typically 1, 2 or 3, carbon-carbon triple bonds. Unless otherwise defined, such alkynyl groups typically contain from 2 to 10 carbon atoms and include, for example, —C₂₋₄alkynyl and —C₃₋₁₀alkynyl. Representative alkynyl groups include, by way of example, ethynyl, n-propynyl, n-but-2-ynyl, n-hex-3-ynyl and the like. The term “alkynylene” means a divalent alkynyl group and includes groups such as —C₂₋₃alkynylene-.

Amino acid residues are often designated as —C(O)—CHR—NH—, where the R moiety is referred to as the “amino acid side chain.” Thus, for the amino acid valine, HO—C(O)—CH[—CH(CH₃)₂]—NH₂, the side chain is —CH(CH₃)₂. Ther term “amino acid side chain” is intended to include side chains of the twenty common naturally occurring amino acids: alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine. Of particular interest are the side chains of non-polar amino acids such as isoleucine, leucine, and valine.

The term “aryl” means a monovalent aromatic hydrocarbon having a single ring (for example, phenyl) or fused rings. Fused ring systems include those that are fully unsaturated (for example, naphthalene) as well as those that are partially unsaturated (for example, 1,2,3,4-tetrahydronaphthalene). Unless otherwise defined, such aryl groups typically contain from 6 to 10 carbon ring atoms and include, for example, —C₆₋₁₀aryl. Representative aryl groups include, by way of example, phenyl and naphthalene-1-yl, naphthalene-2-yl, and the like. The term “arylene” means a divalent aryl group such as phenylene.

The term “cycloalkyl” means a monovalent saturated carbocyclic hydrocarbon group. Unless otherwise defined, such cycloalkyl groups typically contain from 3 to 10 carbon atoms and include, for example, —C₃₋₅cycloalkyl, —C₃₋₆cycloalkyl and —C₃₋₇cycloalkyl. Representative cycloalkyl groups include, by way of example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like. The term “cycloalkylene” means a divalent aryl group such as —C₄₋₈cycloalkylene.

The term “halo” means fluoro, chloro, bromo and iodo.

The term “heteroaryl” means a monovalent aromatic group having a single ring or two fused rings and containing in the ring(s) at least one heteroatom (typically 1 to 3) selected from nitrogen, oxygen or sulfur. Unless otherwise defined, such heteroaryl groups typically contain from 5 to 10 total ring atoms and include, for example, —C₂₋₉heteroaryl. Representative heteroaryl groups include, by way of example, monovalent species of pyrrole, imidazole, thiazole, oxazole, furan, thiophene, triazole, pyrazole, isoxazole, isothiazole, pyridine, pyrazine, pyridazine, pyrimidine, triazine, indole, benzofuran, benzothiophene, benzoimidazole, benzthiazole, quinoline, isoquinoline, quinazoline, quinoxaline and the like, where the point of attachment is at any available carbon or nitrogen ring atom.

The term “optionally substituted” means that group in question may be unsubstituted or it may be substituted one or several times, such as 1 to 3 times or 1 to 5 times. For example, an alkyl group that is “optionally substituted” with 1 to 5 fluoro atoms, may be unsubstituted, or it may contain 1, 2, 3, 4, or 5 fluoro atoms.

The term “protected derivatives thereof” means a derivative of the specified compound in which one or more functional groups of the compound are protected or blocked from undergoing undesired reactions with a protecting or blocking group. Functional groups that may be protected include, by way of example, carboxy groups, amino groups, hydroxyl groups, thiol groups, carbonyl groups and the like. Representative protecting groups for carboxy groups include esters (such as a p-methoxybenzyl ester), amides and hydrazides; for amino groups, carbamates (such as t-butoxycarbonyl) and amides; for hydroxyl groups, ethers and esters; for thiol groups, thioethers and thioesters; for carbonyl groups, acetals and ketals; and the like. Such protecting groups are well-known to those skilled in the art and are described, for example, in T. W. Greene and G. M. Wuts, Protecting Groups in Organic Synthesis, Third Edition, Wiley, New York, 1999, and references cited therein.

All other terms used herein are intended to have their ordinary meaning as understood by those of ordinary skill in the art to which they pertain.

General Synthetic Procedures

Compounds of the invention can be prepared from readily available starting materials using the following general methods, the procedures set forth in the Examples, or by using other methods, reagents, and starting materials that are known to those of ordinary skill in the art. Although the following procedures may illustrate a particular embodiment of the invention, it is understood that other embodiments of the invention can be similarly prepared using the same or similar methods or by using other methods, reagents and starting materials known to those of ordinary skill in the art. It will also be appreciated that where typical or preferred process conditions (for example, reaction temperatures, times, mole ratios of reactants, solvents, pressures, etc.) are given, other process conditions can also be used unless otherwise stated. While optimum reaction conditions will typically vary depending on various reaction parameters such as the particular reactants, solvents and quantities used, those of ordinary skill in the art can readily determine suitable reaction conditions using routine optimization procedures.

Additionally, as will be apparent to those skilled in the art, conventional protecting groups may be necessary or desired to prevent certain functional groups from undergoing undesired reactions. The choice of a suitable protecting group for a particular functional group as well as suitable conditions and reagents for protection and deprotection of such functional groups are well-known in the art. Protecting groups other than those illustrated in the procedures described herein may be used, if desired. For example, numerous protecting groups, and their introduction and removal, are described in T. W. Greene and G. M. Wuts, Protecting Groups in Organic Synthesis, supra. More specifically, the following abbreviations and reagents are used in the schemes presented below:

P¹ represents an “amino-protecting group,” a term used herein to mean a protecting group suitable for preventing undesired reactions at an amino group. Representative amino-protecting groups include, but are not limited to, t-butoxycarbonyl (BOC), trityl (Tr), benzyloxycarbonyl (Cbz), 9-fluorenylmethoxycarbonyl (Fmoc), formyl, trimethylsilyl (TMS), t-butyldimethylsilyl (TBDMS), and the like. Standard deprotection techniques are used to remove the P¹ group. For example, deprotection of the N—BOC groups can use reagents such as TFA in DCM or HCl in 1,4-dioxane, while the Cbz group can be removed by employing catalytic hydrogenation conditions such as H₂ (1 atm), 10% Pd/C in an alcoholic solvent.

P² represents a “carboxy-protecting group,” a term used herein to mean a protecting group suitable for preventing undesired reactions at a carboxy group. Representative carboxy-protecting groups include, but are not limited to, methyl, ethyl, t-butyl, benzyl (Bn), p-methoxybenzyl (PMB), 9-fluoroenylmethyl (Fm), trimethylsilyl (TMS), t-butyldimethylsilyl (TBDMS), diphenylmethyl (benzhydryl, DPM) and the like. Standard deprotection techniques and reagents are used to remove the P² group, and may vary depending upon which group is used. For example, NaOH is commonly used when P² is methyl, an acid such as TFA or HCl is commonly used when P² is t-butyl, and catalytic hydrogenation condition such as H₂ (1 atm) and 10% Pd/C in an alcoholic solvent (“H2/Pd/C”) may also be used when P² is benzyl.

P³ represents a “thiol-protecting group,” a term used herein to mean a protecting group suitable for preventing undesired reactions at a thiol group. Representative thiol-protecting groups include, but are not limited to, ethers, esters such as —C(O)CH₃, and the like. Standard deprotection techniques and reagents such as NaOH, primary alkylamines, and hydrazine, may be used to remove the P³ group.

P⁴ represents a “tetrazole-protecting group,” a term used herein to mean a protecting group suitable for preventing undesired reactions at a tetrazole group. Representative tetrazole-protecting groups include, but are not limited to trityl and diphenylmethyl. Standard deprotection techniques and reagents such as TFA in DCM or HCl in 1,4-dioxane are used to remove the P⁴ group.

P⁵ represents a “hydroxyl-protecting group,” a term used herein to mean a protecting group suitable for preventing undesired reactions at a hydroxyl group. Representative hydroxyl-protecting groups include, but are not limited to alkyl groups such as t-butyl, silyl groups including triC₁₋₆alkylsilyl groups, such as trimethylsilyl (TMS), triethylsilyl (TES), and tert-butyldimethylsilyl (TBDMS); esters (acyl groups) including C₁₋₆alkanoyl groups, such as formyl, acetyl, and pivaloyl, and aromatic acyl groups such as benzoyl; arylmethyl groups such as benzyl (Bn), p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm), and diphenylmethyl (benzhydryl, DPM); and the like. Standard deprotection techniques and reagents are used to remove the P⁵ group, and may vary depending upon which group is used. For example, H₂/Pd/C is commonly used when P⁵ is benzyl, while NaOH is commonly used when P⁵ is an acyl group.

P⁶ represents a “sulfonamide-protecting group,” a term used herein to mean a protecting group suitable for preventing undesired reactions at a sulfonamide group. Representative sulfonamide-protecting groups include, but are not limited to t-butyl and acyl groups. Exemplary acyl groups include aliphatic lower acyl groups such as the formyl, acetyl, phenylacetyl, butyryl, isobutyryl, valeryl, isovaleryl and pivaloyl groups, and aromatic acyl groups such as the benzoyl and 4-acetoxybenzoyl. Standard deprotection techniques and reagents are used to remove the P⁶ group, and may vary depending upon which group is used. For example, HCl is commonly used when P⁶ is t-butyl, while NaOH is commonly used when P⁶ is an acyl group.

P⁷ represents a “phosphate-protecting group or phosphinate-protecting group,” a term used herein to mean a protecting group suitable for preventing undesired reactions at a phosphate or phosphinate group. Representative phosphate and phosphinate protecting groups include, but are not limited to C₁₋₄alkyls, aryl (for example, phenyl) and substituted aryls (for example, chlorophenyl and methylphenyl). The protected group can be represented by —P(O)(OR)₂, where R is a group such as a C₁₋₆alkyl or phenyl. Standard deprotection techniques and reagents such as TMS-I/2,6-lutidine (MeCN), and H₂/Pd/C are used to remove the P⁷ group such as ethyl, and benzyl, respectively.

In addition, L is used to designate a “leaving group,” a term used herein to mean a functional group or atom which can be displaced by another functional group or atom in a substitution reaction, such as a nucleophilic substitution reaction. By way of example, representative leaving groups include chloro, bromo and iodo groups; sulfonic ester groups, such as mesylate, triflate, tosylate, brosylate, nosylate and the like; and acyloxy groups, such as acetoxy, trifluoroacetoxy and the like.

Suitable bases for use in these schemes include, by way of illustration and not limitation, potassium carbonate, calcium carbonate, sodium carbonate, triethylamine, pyridine, 1,8-diazabicyclo-[5.4.0]undec-7-ene (DBU), N,N-diisopropylethylamine (DIPEA), sodium hydroxide, potassium hydroxide, potassium t-butoxide, and metal hydrides.

Suitable inert diluents or solvents for use in these schemes include, by way of illustration and not limitation, tetrahydrofuran (THF), acetonitrile (MeCN), N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), toluene, dichloromethane (DCM), chloroform (CHCl₃), carbon tetrachloride (CCl₄), 1,4-dioxane, methanol, ethanol, water, and the like.

Suitable carboxylic acid/amine coupling reagents include benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate (BOP), benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate (PyBOP), O-(7-azabenzotriazol-1-yl-N,N,N′,N′ tetramethyluronium hexafluorophosphate (HATU), N,N′-dicyclohexylcarbodiimide (DCC), N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC), carbonyldiimidazole (CDI), and the like. Coupling reactions are conducted in an inert diluent in the presence of a base, and are performed under conventional amide bond-forming conditions.

All reactions are typically conducted at a temperature within the range of about −78° C. to 100° C., for example at room temperature. Reactions may be monitored by use of thin layer chromatography (TLC), high performance liquid chromatography (HPLC), and/or LCMS until completion. Reactions may be complete in minutes, or may take hours, typically from 1-2 hours and up to 48 hours. Upon completion, the resulting mixture or reaction product may be further treated in order to obtain the desired product. For example, the resulting mixture or reaction product may be subjected to one or more of the following procedures: concentrating or partitioning (for example, between EtOAc and water or between 5% THF in EtOAc and 1M phosphoric acid); extraction (for example, with EtOAc, CHCl₃, DCM, HCl); washing (for example, with saturated aqueous NaCl, saturated NaHCO₃, Na₂CO₃ (5%), CHCl₃ or 1M NaOH); drying (for example, over MgSO₄, over Na₂SO₄, or in vacuo); filtering; crystallizing (for example, from EtOAc and hexane); and/or being concentrated (for example, in vacuo).

By way of illustration, compounds of formula I, as well as their salts, solvates, and prodrugs can be prepared by one or more of the following exemplary processes.

Scheme I Peptide Coupling Reaction and Optional Deprotection

The X moiety contains one or more amide groups, and therefore the compounds of the invention are typically formed by a coupling reaction under conventional amide bond-forming conditions, followed by a deprotection step if needed. In Scheme I, the A and B moieties couple to form X, and the sum of a and b is in the range of 0 to 11. Thus, one moiety comprises an amine group and one moiety comprises a carboxylic acid group, i.e., A is —NH₂ and B is —COOH or A is —COOH and B is —NH₂.

For example, to synthesize a compound of formula I where X is —CONH—, A would be —COOH and B would be —NH₂. Similarly, A as —NH₂ and B as —COOH would couple to form —NHCO— as the X moiety. A and B can be readily modified if a longer X is desired, whether it contains an alkylene portion or additional amide groups. For example, A as —CH₂NH₂ and B as —COOH would couple to form —CH₂NHCO— as the X moiety.

It is understood that the carbon atoms in the —(CH₂)_(a) and —(CH₂)_(b) groups make up the “X” linker. Therefore, these carbon atoms may be substituted with one or more Rob groups. Furthermore, one —CH₂— group in the —(CH₂)_(a) or the —(CH₂)_(b) group may be replaced with a —C₃₋₈cycloalkylene-, —CR^(4d)═CH—, or —CH═CR^(4d)— group.

Ar* represents Ar—R¹*, where R¹* may represent R¹ as defined herein, or a protected form of R¹ (e.g., -tetrazol-5-yl-P⁴ or —C(O)O—P² such as —C(O)O—C₁₋₆alkyl), or a precursor of R¹ (e.g., —CN that is then converted to tetrazole, or nitro that is then converted to amino from which the desired R¹ is prepared). R⁵* represents R⁵ as defined herein, or a protected form of R⁵. Therefore, when R¹* represents R¹ and R⁵* represents R⁵, the reaction is complete after the coupling step.

On the other hand, when R¹* represents a protected form of R¹ and/or R⁵* represents a protected form of R⁵, a subsequent global or sequential deprotection step would yield the non-protected compound. Similarly, when R¹* represents a precursor of R′, a subsequent conversion step would yield the desired compound. Reagents and conditions for the deprotection vary with the nature of protecting groups in the compound. Typical deprotection conditions when R⁵* represents C₀₋₃alkylene-S—P³, include treating the compound with NaOH in an alcoholic solvent at 0° C. or room temperature to yield the non-protected compound. Typical deprotection conditions when R¹* represents C(O)O—P² where P² refers to t-butyl include treating the compound with TFA in DCM at room temperature to yield the non-protected compound. Thus, one method of preparing compounds of the invention involves coupling compound (1) and (2), with an optional deprotection step when R¹* is a protected form of R¹ and/or R⁵* is a protected form of R⁵, thus forming a compound of formula I or a pharmaceutically acceptable salt thereof.

Examples of compound (1) include: 3-(2′-t-butoxycarbonylbiphenyl-4-ylmethyl)-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic acid; and 4-(6-aminomethyl-4-methyl-2-propylbenzoimidazol-1-ylmethyl)-2-fluorobenzoic acid methyl ester. Examples of compound (2) include: (R)-1-benzyl-2-hydroxycarbamoylethyl)carbamic acid; (R)-2-((R)-2-amino-3-phenylpropyldisulfanyl)-1-benzylethylamine; (S)-2-acetylsulfanyl-4-methylpentanoic acid; 2-acetylsulfanylmethyl-4-methylpentanoic acid; (S)-2-acetylsulfanylmethyl-4-methylpentanoic acid; and (R)-2-(2-benzyloxycarbamoyl-3-phenypropionyl-amino)succinic acid 1-methyl ester.

Compound (1) Esterification of the Benzoimidazole Carboxylic Acid

An acidic solution (e.g., HCl:water, 30:70) is added to compound (1a) dissolved in a solvent such as methanol. The mixture is refluxed until completion (˜12-36 hours), then concentrated under reduced pressure or in vacuo to afford compound (1b) as a solid. The recovered material may be used without further processing. Alternately, the solid may be dissolved in ethyl acetate, washed with saturated NaHCO₃, dried over MgSO₄ and concentrated. Acidic solutions used include sulfuric acid and a hydrogen chloride solution (30:70, HCl:H₂O). Examples of compound (1a) include 7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic acid (R²=methyl, R³=propyl), which is commercially available, and is particularly well-suited for use in preparing compounds of the invention. If desired, different R² and R³ groups can be introduced in later steps. Examples of compound (1b) include 7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic acid methyl ester.

Conversion of the Benzoimidazole Ester to an Alcohol

LiAlH₄ in a solvent such as THF is cooled to 0° C. and stirred under nitrogen. A solution of compound (1b) in the same solvent is added dropwise and the mixture is stirred at 0° C. After 1 hour, the mixture is warmed to room temperature and stirred until completion (˜12 hours). The mixture is then cooled to 0° C. Ethyl acetate is added to quench any remaining LiAlH₄. The reaction is then quenched with saturated NH₄Cl. The mixture is washed with ethyl acetate and the layers separated. The organic layer is then dried over MgSO₄ and concentrated. The recovered solid is flash chromatographed to produce compound (1c). Examples of compound (1c) include (7-methyl-2-propyl-3H-benzoimidazol-5-yl)methanol.

Preparation of Aryl Compound

Compound (1d) can be prepared using synthetic methods that are reported in the literature, for example Duncia et al. (1991) J. Org. Chem. 56: 2395-400, and references cited therein. Alternatively, the starting material in a protected form (1e) may be commercially available. Using a commercially available non-protected starting material (1d), the R¹ group is first protected to form protected intermediate (1e), then the leaving group (L) is added to form compound (10, for example, by a halogenation reaction. For example, a bromination reaction of a methyl group of N-triphenylmethyl-5-[4′-methylbiphenyl-2-yl]tetrazole is described in Chao et al. (2005) J. Chinese Chem. Soc. 52:539-544. In addition, when Ar* has a —CN group, it can be subsequently converted to the desired tetrazolyl group, which may be protected. Conversion of the nitrile group is readily achieved by reaction with a suitable azide such as sodium azide, trialkyltin azide (particularly tributyltin azide) or triaryltin azide. Compound (10 when Ar has one of the remaining formulas is readily synthesized using similar techniques or other methods as are well known in the art.

Exemplary methods of preparing compound (1f) include the following. A solution of the starting material (1d) and thionyl chloride are stirred at room temperature. After completion of the reaction, the mixture is concentrated in vacuo to afford a solid, which is dissolved in a solvent such as THF and cooled to 0° C. Potassium t-butoxide is then added. Upon completion of the reaction, the mixture is partitioned between ethyl acetate and water. The organic layer is washed with saturated aqueous NaCl, dried over MgSO₄, filtered, and concentrated to afford compound (1e). Alternately, HCl is added to a solution of the starting material (1d) and a solvent such as methanol. The mixture is heated to reflux and stirred until the reaction is complete (˜48 hours), then cooled and concentrated. The recovered material is dried in vacuo to obtain compound (1e). Intermediate (1e), benzoyl peroxide, and N-bromosuccinimide are dissolved in CCl₄ or benzene, and heated to reflux. The mixture is stirred to completion, cooled to room temperature, filtered, and concentrated in vacuo. The resulting residue is crystallized from diethyl ether and hexane or flash chromatographed to give compound (1f).

Examples of compound (id) include: 4′-methylbiphenyl-2-carboxylic acid; 2,3-difluoro-4-methyl-benzoic acid; and 2-fluoro-4-methylbenzoic acid. Examples of compound (1e) include N-triphenylmethyl-5-[4′-methylbiphenyl-2-yl]tetrazole. Examples of compound (1f) include: 4′-bromomethylbiphenyl-2-carboxylic acid t-butyl ester; 4-bromomethyl-2-fluorobenzoic acid methyl ester; 5-(4′-bromomethylbiphenyl-2-yl)-1-trityl-1H-tetrazole; 4-bromomethylbenzoic acid methyl ester; 4-bromomethyl-2,3-difluoro-benzoic acid methyl ester; 4′-formyl-biphenyl-2-sulfonic acid t-butylamide; 4′-amino-methylbiphenyl-2-carboxylic acid t-butyl ester; and 4′-bromomethyl-3′-fluorobiphenyl-2-carboxylic acid t-butyl ester.

Compound (1f) where R¹ is —SO₂NHR^(1d) may be synthesized as follows:

The starting material, 2-bromobenzene-1-sulfonamide, is commercially available. Reaction of 2-bromobenzene-1-sulfonamide in a solvent such as DMF, with 1,1-dimethoxy-N,N-dimethylmethanamine, followed by the addition of sodium hydrogen sulfate in water, yields 2-bromo-N-[1-dimethylaminometh-(E)-ylidene]benzenesulfonamide. This compound is reacted with 4-methylphenylboronic acid to yield 4′-methylbiphenyl-2-sulfonic acid 1-dimethylaminometh-(E)-ylideneamide, then the —(CH₂)_(r)—L¹ moiety is added, for example, by a halogenation reaction, to form compound (1f).

Compound (1f) where the Ar moiety is substituted may be synthesized as follows:

The starting material, 2-bromobenzoic acid, is commercially available. Reaction of 2-bromobenzoic acid in a suitable solvent, with t-butyl alcohol, DCC and DMAP, yields 2-bromo-benzoic acid t-butyl ester. This compound is reacted with 3-fluoro-4-methylphenylboronic acid to yield 3′-fluoro-4′-methylbiphenyl-2-carboxylic acid t-butyl ester. The —(CH₂)_(r)—L¹ moiety is then added to form compound (1f), for example, by a halogenation reaction.

Alkylation Procedure

Compound (1g) is formed by the following alkylation reaction. A solution of compound (1b) in a solvent such as DMF is cooled in an ice bath. Sodium hydride (60 wt % in mineral oil) is added, followed by the addition of compound (1f). The mixture is gradually warmed to room temperature, then stirred until completion (up to 12 hours). The mixture is then cooled and concentrated in vacuo, and the resultant residue partitioned between saturated aqueous NaCl or LiCl and ethyl acetate. The organic layer is collected, dried over MgSO₄ and evaporated to dryness, yielding compound (1g).

Hydrolysis Procedure Carboxylic Acid-Terminal A Group

Compound (10 and a solvent such as THF are combined with 1 M NaOH in water, and the mixture is stirred at room temperature until completion (˜12 hours). The mixture is then acidified to a pH of 4, for example with 1M HCl. The solution is extracted with ethyl acetate, and concentrated under reduced pressure to afford compound (1). Examples of compound (1g′) include 3-(2′-t-butoxycarbonylbiphenyl-4-ylmethyl)-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic acid methyl ester.

Alcohol to Amine Conversion Amine-Terminal A Group

Compound (1g″) is combined with a solvent such as DCM, a base such as DIPEA, and 4-dimethylaminopyridine. Methanesulfonyl chloride is added at 0° C., and the mixture is warmed to room temperature. After the reaction is complete (˜3 hours), the mixture is concentrated and the material redissolved in solvent. Sodium azide is added and the mixture is heated at 80° C. for 1 hour, then cooled to room temperature. The solution is partitioned between ethyl acetate and 10% LiCl (aqueous). The organic layer is concentrated and flash chromatographed. The product is dissolved in a solvent such as THF. Triphenylphosphine is added and the mixture is stirred at room temperature for 30 minutes. Water is added and the mixture heated to 60° C. After the reaction is complete (˜3 hours), the mixture is cooled to room temperature and concentrated in vacuo. The concentrate is dissolved in ethyl acetate, and washed in 1N HCl. The aqueous layer is extracted with ethyl acetate then basified to pH-10, for example with 6N NaOH. The solution is saturated with NaCl, extracted with a solvent such as DCM, and dried over MgSO₄. The organic layer is then concentrated to afford compound (1). Examples of compound (1g′″) include 2-fluoro-4-(6-hydroxymethyl-4-methyl-2-propyl-benzoimidazol-1-ylmethyl)benzoic acid methyl ester.

Compound (2)

Compound (2) can be readily synthesized by following techniques described in the literature, for example, Neustadt et al (1994) J. Med. Chem. 37:2461-2476 and Moree et al. (1995) J. Org. Chem. 60: 5157-69, as well as by using the exemplary procedures described below. Examples of compound (2), depicted without chirality, include:

Since compound (2) has a chiral center, it may be desirable to synthesize a particular stereoisomer, and examples are provided as follows.

Preparation of Chiral Amino Hydroxamate Compound (2^(i))

A base such as DIPEA and a coupling agent such as EDC are added to a solution of compound (2a) in DMF containing HOBt and hydroxylamine hydrochloride. The mixture is stirred at room temperature until completion (˜12 hours), then concentrated in vacuo. The resulting material is distributed between 5% THF in ethyl acetate and 1M phosphoric acid. The organic layer is collected and washed with a base such as 1M NaOH. The alkaline aqueous layer is then acidified, for example with 1M phosphoric acid, and extracted with ethyl acetate. The organic layer is evaporated and the residue purified by silica gel chromatography to afford compound (2′). Examples of compound (2a) include (R)-3-t-butoxycarbonylamino-4-phenylbutyric acid.

Preparation of Chiral Amino Sulfhydryl Dimer Compound (2^(ii))

Diisopropyl azodicarboxylate is added to a solution of triphenylphosphine in a solvent such as THF, cooled in an ice bath. The solution is stirred and compound (2b) and thioacetic acid are added. The mixture is stirred at 0° C. for 1 hour, then stirred at room temperature until completion (˜12 hours). The mixture is stripped, diluted with ethyl acetate, and washed with a cold saturated NaHCO₃ solution. The organic layer is dried over MgSO₄, and the filtrate evaporated to dryness. The resulting material is flash chromatographed to provide compound (2c). Compound (2c) is dissolved in solvent, followed by the addition of a base such as 1M LiOH. Air is bubbled through the solution for 1 hour followed by the addition of solvent. The mixture is stirred at room temperature until completion (˜24 hours). The solution is then acidified to pH-5, for example with acetic acid. The precipitate is filtered and rinsed with deionized water, producing the compound (2d) dimer. The solid is suspended in MeCN, then concentrated under reduced pressure. The recovered material is dissolved in 4M HCl in 1,4-dioxane and stirred at room temperature until the reaction is complete (˜2 hours). The mixture is then concentrated under reduced pressure, and triturated with ethyl acetate. The product is filtered, washed with ethyl acetate, and dried in vacuo to provide compound (2¹¹). Examples of compound (2b) include ((R)-1-benzyl-2-hydroxyethyl)carbamic acid t-butyl ester.

Preparation of Chiral Sulfanyl Acid Compound (2^(iii))

Compound (2e) is formed by dissolving a compound such as D-leucine (for R⁶=isobutyl, for example) in 3M HBr (aqueous) and cooled to 0° C. A solution of sodium nitrite in water is added, and the mixture stirred at 0° C. until completion (2.5 hours). The mixture is then extracted with ethyl acetate, washed with saturated aqueous NaCl, dried over MgSO₄, filtered, and concentrated to afford compound (2e). Compound (2e) is combined with potassium thioacetate or sodium thioacetate and DMF, and the mixture stirred at room temperature until completion (1 hour). Water is added. The mixture is then extracted with ethyl acetate, washed with saturated aqueous NaCl, dried over MgSO₄, filtered, and concentrated to provide compound (2^(iii)). The product is purified by silica gel chromatography. Examples of compound (2e) include (R)-2-bromo-4-methylpentanoic acid. Examples of compound (2¹′) include (S)-2-acetylsulfanyl-4-methylpentanoic acid.

Preparation of Sulfanyl Acid Compound (2′)

Compound (20 is mixed with diethylamine and cooled in an ice bath. An aqueous formaldehyde solution (37%) is then added, and the mixture stirred at 0° C. for approximately 2 hours, warmed to room temperature and stirred overnight. The mixture is then extracted with ether, washed, dried, and evaporated to dryness, to provide compound (2g). Compound (2g) is then dissolved in 1,4-dioxane, and a 1M NaOH solution is added. The mixture is stirred at room temperature until completion (approximately 2 days). The organic solvent is removed in vacuo, and the aqueous residue is rinsed with EtOAc and acidified to approximately pH 1 with concentrated HCl. The product is extracted with EtOAc, dried, and evaporated to dryness to yield compound (2h). Compound (2h) is combined with thiolacetic acid (10 mL), and the mixture is stirred at 80° C. until completion (approximately 2 hours), then concentrated to dryness to yield compound (r), which is dissolved in toluene and concentrated to remove any trace of thiolacetic acid. Examples of compound (20 include 2-benzylmalonic acid monoethyl ester (R⁶=benzyl) and 2-isobutylmalonic acid monoethyl ester (R⁶=isobutyl).

Preparation of Chiral Sulfanyl Acid Compound (2′)

Compound (2i), (S)-4-benzyl-2-oxazolidinone, is commercially available. Compound (2j) is also typically commercially available or can be readily synthesized. For example, R⁶—CH₂—COOH (for example, isocaproic acid or 3-phenylpropionic acid) is dissolved in methylene chloride and thionyl chloride is added. The mixture is stirred at room temperature until the reaction is complete (for example, overnight), and then concentrated to provide (2j). Examples of compound (2j) include 4-methylpentanoyl chloride and 3-phenylpropionyl chloride.

Compound (2i) is dissolved in a suitable solvent and cooled (˜78° C.) under nitrogen. n-Butyllithium in hexanes is added dropwise and stirred, followed by the addition of (2j) dropwise. The mixture is stirred at −78° C., then warmed to 0° C. Saturated NaHCO₃ is added and the mixture warmed to room temperature. The mixture is extracted, washed, dried, filtered, and concentrated to afford (2k). Compound (2k) is dissolved in DCM and stirred at 0° C. under nitrogen. 1M Titanium tetrachloride is added, followed by 1,3,5-trioxane, all in appropriate solvents. A second equivalent of 1M titanium tetrachloride is added and the mixture stirred at 0° C. until the reaction is complete. The reaction is then quenched with saturated ammonium chloride. Appropriate solvents are added, the aqueous phase is extracted, and the organic layers are combined, dried, filtered, and concentrated to provide (21), which can then be purified by silica gel chromatography or used in the next step without further purification. Compound (21) is dissolved in a solvent, to which is added 9 M hydrogen peroxide in water, followed by the dropwise addition of 1.5 M lithium hydroxide monohydrate in water. The mixture is warmed to room temperature and stirred. Optionally, potassium hydroxide may be added and the mixture heated at 60° C. then cooled at room temperature. To this is added an aqueous solution of sodium sulfite followed by water and chloroform. The aqueous layer is extracted, acidified and extracted again. The organic layer is washed, dried, filtered, and rotovaped to provide (2m). Triphenylphosphine is dissolved in an appropriate solvent and cooled at 0° C. (ice bath). Diisopropyl azodicarboxylate is added dropwise and the mixture stirred. Compound (2m) and thioacetic acid, dissolved in an appropriate solvent, are added dropwise to the mixture. After the addition, the mixture is removed from the ice bath and stirred at room temperature until the reaction is complete (approximately 3.5 hours), concentrated, and then partitioned. The organic layer is extracted and the combined aqueous extracts washed, acidified and extracted. The organic layer is washed again, dried, filtered, and rotovaped to provide compound (2^(v)). Examples of compound (2^(v)) include (S)-2-acetylsulfanylmethyl-4-methylpentanoic acid.

Scheme II Peptide Coupling Reaction

Compounds of formula I can also be prepared by coupling compound (3) and compound (1) where A is —COOH, followed by reaction with compound (4). In compounds (1) and (3), the sum of a and b is in the range of 0 to 11. In compounds (3) and (4), the [R⁵] and [[R⁵]] moieties represent portions of the R⁵ moiety. For example, if R⁵ is —CH₂C(O)N(OH)H, then [R⁵] would be —CH₂C(O)— and [[R⁵]] would be —N(OH)H.

The amide coupling of compounds (1) and (3) is typically conducted at room temperature with coupling reagents such as pentafluorophenol combined with EDC and DIPEA in solvents such as DMF. Compounds (3) and (4) are available commercially or can be readily synthesized by techniques that are well known in the art. Examples of compound (3) include 3-amino-4-(3-chlorophenyl)butyric acid. Examples of compound (4) include H₂NOH, H₂NO-benzyl, and H₂NO-t-butyl.

Scheme III Peptide Coupling Reaction

Compounds of formula I can also be prepared by coupling compounds (1) and (5) under conventional amide bond-forming conditions. This synthesis is particularly useful for preparing compounds of formula I where more than one —CH₂— moiety in the alkylene is replaced with —NR^(4a)—C(O)— or —C(O)—NR^(4a)—. In the compounds depicted below, b will typically be at least 1 and the sum of a, b and c is in the range of 1 to 10.

Compound (5) is formed by converting the ester compound (6) to the acid with a base, converting the acid to the dioxopyrrolidinyl ester with N-hydroxysuccinimide, then converting the dioxopyrrolidinyl ester with compound (7) to form compound (5). Compound (5) may have one or more R^(4b) substituents on the carbon atoms in the —(CH₂)_(b)— and/or —(CH₂)_(c)— portions.

Compound (6) can be readily synthesized by following the techniques described in the literature, for example, Fournie-Zaluski et al. (1985) J. Med. Chem. 28(9):1158-1169). Examples of compound (6) include 2-benzyl-N-benzyloxymalonamic acid ethyl ester. Compound (7) is available commercially or can be readily synthesized by techniques that are well known in the art. This reactant is particularly useful where a carbon atom in the alkylene moiety in X is substituted with an R^(4b) group. For example, 2-aminosuccinic acid 1-methyl ester is an example of compound (7) that is useful for preparing compounds where R^(4b) is —COOH, where the R^(4b) moiety is in a protected form, —C(O)OCH₃.

Scheme IV Two-Step Reaction

Compounds of formula I can also be prepared by a two-step method, where the first step involves the addition of a halogen-substituted alkanoic acid (“L-acid”) such as α-bromoisocaproic acid to compound (1), to provide Intermediate (8), where L is a leaving group such as bromo. Intermediate (8) is then reacted with a thiol or sulfur-containing nucleophilic reactant that contains the desired R⁵* group, for example, potassium thioacetate or thiourea.

Scheme V Prodrug Synthesis

Prodrugs can be readily synthesized using the techniques described above. In addition, prodrugs can be formed by further modifying active compounds of formula I where Ar**—COOH represents Ar—R′ and R⁵ is —C₀₋₃alkylene-SH, as shown below:

Thus, both prodrug A and prodrug C can be readily synthesized from the corresponding active compound.

If desired, pharmaceutically acceptable salts of the compounds of formula I can be prepared by contacting the free acid or base form of a compound of formula I with a pharmaceutically acceptable base or acid.

Certain intermediates described herein are believed to be novel and accordingly, such compounds are provided as further aspects of the invention including, for example, the compounds of formulas II, III and IV, and salts thereof:

where Ar* is Ar—R¹*; Ar, r, n, and R²⁻³, X, and R⁵⁻⁷ are as defined for formula I; and R¹* is —C(O)O—P², —SO₂O—P⁵, —SO₂NH—P⁶, —P(O)(O—P⁷)₂, —OCH(CH₃)—C(O)O—P², —OCH(aryl)-C(O)O—P², or tetrazol-5-yl-P⁴; where P² is a carboxy-protecting group, P⁴ is a tetrazole-protecting group, P⁵ is a hydroxyl-protecting group, P⁶ is a sulfonamide-protecting group, and P⁷ is a phosphate-protecting group or phosphinate-protecting group;

where Ar, r, n, R²⁻³, X, and R⁶⁻⁷ are as defined for formula I; R⁵* is selected from —C₀₋₃alkylene-S—P³, —C₀₋₃alkylene—C(O)NH(O—P⁵), —C₀₋₃alkylene-N(O—P⁵)—C(O)R^(5d), —C₀₋₁alkylene-NHC(O)CH₂S—P³, —NH—C₀₋₁alkylene-P(O)(O—P⁷)₂, —C₀₋₃alkylene-P(O)(O—P⁷)—R^(5f), —C₀₋₂alkylene-CHR^(5g)—C(O)O—P² and —C₀₋₃alkylene—C(O)NR^(5l)—CHR^(5l)—C(O)O—P²; and R^(5d-1) are as defined for formula I; where P² is a carboxy-protecting group, P³ is a thiol-protecting group, P⁵ is a hydroxyl-protecting group, and P⁷ is a phosphate-protecting group or phosphinate-protecting group; and

where Ar* is Ar—R¹*; Ar, r, n, R²⁻³, X, and R⁶⁻⁷ are as defined for formula I; R¹* is —C(O)O—P², —SO₂O—P⁵, —SO₂NH—P⁶, —P(O)(O—P⁷)₂, —OCH(CH₃)—C(O)O—P², —OCH(aryl)-C(O)O—P², or tetrazol-5-yl-P⁴; R⁵* is —C₀₋₃alkylene-S—P³, —C₀₋₃alkylene—C(O)NH(O—P⁵), —C₀₋₃alkylene-N(O—P⁵)—C(O)R^(5d), —C₀₋₁alkylene-NHC(O)CH₂S—P³, —NH—C₀₋₁alkylene-P(O)(O—P⁷)₂, —C₀₋₃alkylene-P(O)(O—P⁷)—R^(5f), —C₀₋₂alkylene-CHR^(5g)—C(O)O—P², or —C₀₋₃alkylene—C(O)NR^(5h)—CHR^(5l)—C(O)O—P²; and R^(5d-1) are as defined for formula I; where P² is a carboxy-protecting group, P³ is a thiol-protecting group, P⁴ is a tetrazole-protecting group, P⁵ is a hydroxyl-protecting group, P⁶ is a sulfonamide-protecting group, and P⁷ is a phosphate-protecting group or phosphinate-protecting group. Thus, another method of preparing compounds of the invention involves deprotecting a compound of formula II, III, or IV.

In one embodiment, r and n are both 1, and the intermediates are compounds of formulas IIa, IIIa and IVa, and salts thereof:

where Ar*, Ar, R²⁻³, X, R⁵*, R⁵⁻⁷ are as defined above.

Further details regarding specific reaction conditions and other procedures for preparing representative compounds of the invention or intermediates thereof are described in the Examples set forth below.

Utility

Compounds of the invention possess angiotensin II type 1 (AT₁) receptor antagonist activity. In one embodiment, compounds of the invention are selective for inhibition of the AT₁ receptor over the AT₂ receptor. Compounds of the invention also possess neprilysin (NEP) inhibition activity, that is, the compounds are able to inhibit enzyme-substrate activity. In another embodiment, the compounds do not exhibit significant inhibitory activity at the angiotensin-converting enzyme. Compounds of formula I may be active drugs as well as prodrugs. Thus, when discussing the activity of compounds of the invention, it is understood that any such prodrugs have the expected activity once metabolized.

One measure of the affinity of a compound for the AT₁ receptor is the inhibitory constant (K_(i)) for binding to the AT₁ receptor. The pK_(i) value is the negative logarithm to base 10 of the K_(i). One measure of the ability of a compound to inhibit NEP activity is the inhibitory concentration (IC₅₀), which is the concentration of compound that results in half-maximal inhibition of substrate conversion by the NEP enzyme. The pIC₅₀ value is the negative logarithm to base 10 of the IC₅₀. Compounds of formula I and pharmaceutically acceptable salts thereof that have both AT₁ receptor-antagonizing activity and NEP enzyme-inhibiting activity are of particular interest, including those that exhibit a pK_(i) at the AT₁ receptor greater than or equal to about 5.0, and exhibit a pIC₅₀ for NEP greater than or equal to about 5.0.

In one embodiment, compounds of interest have a pK_(i) at the AT₁ receptor ≧about 6.0, a pK_(i) at the AT₁ receptor ≧about 7.0, or a pK_(i) at the AT₁ receptor ≧about 8.0. Compounds of interest also include those having a plC₅₀ for NEP ≧about 6.0 or a plC₅₀ for NEP ≧about 7.0. In another embodiment, compounds of interest have a pK_(i) at the AT₁ receptor within the range of about 8.0-10.0 and a plC₅₀ for NEP within the range of about 7.0-10.0.

In another embodiment, compounds of particular interest have a plc for binding to an AT₁ receptor greater than or equal to about 7.5 and a NEP enzyme plC₅₀ greater than or equal to about 7.0. In another embodiment, compounds of interest have a pK_(i) greater than or equal to about 8.0 and a plC₅₀ greater than or equal to about 8.0.

It is noted that in some cases, compounds of the invention, while still having dual activity, may possess either weak AT₁ receptor antagonist activity or weak NEP inhibition activity. In such cases, those of skill in the art will recognize that these compounds still have utility as primarily either a NEP inhibitor or an AT₁ receptor antagonist, respectively, or have utility as research tools.

Exemplary assays to determine properties of compounds of the invention, such as the AT₁ receptor binding and/or NEP inhibiting activity, are described in the Examples and include by way of illustration and not limitation, assays that measure AT₁ and AT₂ binding (described in Assay 1), and NEP inhibition (described in Assay 2). Useful secondary assays include assays to measure ACE inhibition (also described in Assay 2) and aminopeptidase P (APP) inhibition (described in Sulpizio et al. (2005) JPET 315:1306-1313). A pharmacodynamic assay to assess the in vivo inhibitory potencies for ACE, AT₁, and NEP in anesthetized rats is described in Assay 3 (see also Seymour et al. Hypertension 7(Suppl I):I-35-1-42, 1985 and Wigle et al. Can. J. Physiol. Pharmacol. 70:1525-1528, 1992), where AT₁ inhibition is measured as the percent inhibition of the angiotensin II pressor response, ACE inhibition is measured as the percent inhibition of the angiotensin I pressor response, and NEP inhibition is measured as increased urinary cyclic guanosine 3′,5′-monophosphate (cGMP) output. Useful in vivo assays include the conscious spontaneously hypertensive rat (SHR) model, which is a renin dependent hypertension model useful for measuring AT₁ receptor blocking (described in Assay 4; see also Intengan et al. (1999) Circulation 100(22):2267-2275 and Badyal et al. (2003) Indian Journal of Pharmacology 35:349-362), and the conscious desoxycorticosterone acetate-salt (DOCA-salt) rat model, which is a volume dependent hypertension model useful for measuring NEP activity (described in Assay 5; see also Trapani et al. (1989) J. Cardiovasc. Pharmacol. 14:419-424, Intengan et al. (1999) Hypertension 34(4):907-913, and Badyal et al. (2003) supra). Both the SHR and DOCA-salt models are useful for evaluating the ability of a test compound to reduce blood pressure. The DOCA-salt model is also useful to measure a test compound's ability to prevent or delay a rise in blood pressure. Compounds of the invention are expected to antagonize the AT₁ receptor and/or inhibit the NEP enzyme in any of the assays listed above, or assays of a similar nature. Thus, the aforementioned assays are useful in determining the therapeutic utility of compounds of the invention, for example, their utility as antihypertensive agents. Other properties and utilities of compounds of the invention can be demonstrated using other in vitro and in vivo assays well-known to those skilled in the art.

Compounds of the invention are expected to be useful for the treatment and/or prevention of medical conditions responsive to AT₁ receptor antagonism and/or NEP inhibition. Thus it is expected that patients suffering from a disease or disorder that is treated by antagonizing the AT₁ receptor and/or by inhibiting the NEP enzyme can be treated by administering a therapeutically effective amount of a compound of the invention. For example, by antagonizing the AT₁ receptor and thus interfering with the action of angiotensin II on its receptors, these compounds are expected to find utility in preventing the increase in blood pressure produced by angiotensin II, a potent vasopressor. In addition, by inhibiting NEP, the compounds are also expected to potentiate the biological effects of endogenous peptides that are metabolized by NEP, such as the natriuretic peptides, bombesin, bradykinins, calcitonin, endothelins, enkephalins, neurotensin, substance P and vasoactive intestinal peptide. For example, by potentiating the effects of the natriuretic peptides, compounds of the invention are expected to be useful to treat glaucoma. The compounds are also expected to have other physiological actions, for example, on the renal, central nervous, reproductive and gastrointestinal systems.

Compounds of the invention are expected to find utility in treating and/or preventing medical conditions such as cardiovascular and renal diseases. Cardiovascular diseases of particular interest include heart failure such as congestive heart failure, acute heart failure, chronic heart failure, and acute and chronic decompensated heart failure. Renal diseases of particular interest include diabetic nephropathy and chronic kidney disease. One embodiment of the invention is directed to a method for treating hypertension, comprising administering to a patient a therapeutically effective amount of a compound of the invention. Typically, the therapeutically effective amount is the amount that is sufficient to lower the patient's blood pressure. In one embodiment, the compound is administered as an oral dosage form.

Another embodiment of the invention is directed to a method for treating heart failure, comprising administering to a patient a therapeutically effective amount of a compound of the invention. Typically, the therapeutically effective amount is the amount that is sufficient to lower blood pressure and/or improve renal functions. In one embodiment, the compound is administered as an intravenous dosage form. When used to treat heart failure, the compound may be administered in combination with other therapeutic agents such as diuretics, natriuretic peptides, and adenosine receptor antagonists.

Compounds of the invention are also expected to be useful in preventative therapy, for example in preventing the progression of cardiac insufficiency after myocardial infarction, preventing arterial restenosis after angioplasty, preventing thickening of blood vessel walls after vascular operations, preventing atherosclerosis, and preventing diabetic angiopathy.

In addition, as NEP inhibitors, compounds of the invention are expected to inhibit enkephalinase, which will inhibit the degradation of endogenous enkephalins. Thus, such compounds may also find utility as analgesics. Due to their NEP inhibition properties, compounds of the invention are also expected to be useful as antitussive agents and antidiarrheal agents (for example, for the treatment of watery diarrhea), as well as find utility in the treatment of menstrual disorders, preterm labor, pre-eclampsia, endometriosis, reproductive disorders (for example, male and female infertility, polycystic ovarian syndrome, implantation failure), and male and female sexual dysfunction, including male erectile dysfunction and female sexual arousal disorder. More specifically, the compounds of the invention are expected to be useful in treating female sexual dysfunction, which is often defined as a female patient's difficulty or inability to find satisfaction in sexual expression. This covers a variety of diverse female sexual disorders including, by way of illustration and not limitation, hypoactive sexual desire disorder, sexual arousal disorder, orgasmic disorder and sexual pain disorders. When used to treat such disorders, especially female sexual dysfunction, compounds of the invention may be combined with one or more of the following secondary agents: PDE5 inhibitors, dopamine agonists, estrogen receptor agonists and/or antagonists, androgens, and estrogens.

The amount of the compound of the invention administered per dose or the total amount administered per day may be predetermined or it may be determined on an individual patient basis by taking into consideration numerous factors, including the nature and severity of the patient's condition, the condition being treated, the age, weight, and general health of the patient, the tolerance of the patient to the active agent, the route of administration, pharmacological considerations such as the activity, efficacy, pharmacokinetics and toxicology profiles of the compound and any secondary agents being administered, and the like. Treatment of a patient suffering from a disease or medical condition (such as hypertension) can begin with a predetermined dosage or a dosage determined by the treating physician, and will continue for a period of time necessary to prevent, ameliorate, suppress, or alleviate the symptoms of the disease or medical condition. Patients undergoing such treatment will typically be monitored on a routine basis to determine the effectiveness of therapy. For example, in treating hypertension, blood pressure measurements may be used to determine the effectiveness of treatment. Similar indicators for other diseases and conditions described herein, are well-known and are readily available to the treating physician. Continuous monitoring by the physician will insure that the optimal amount of the compound of the invention will be administered at any given time, as well as facilitating the determination of the duration of treatment. This is of particular value when secondary agents are also being administered, as their selection, dosage, and duration of therapy may also require adjustment. In this way, the treatment regimen and dosing schedule can be adjusted over the course of therapy so that the lowest amount of active agent that exhibits the desired effectiveness is administered and, further, that administration is continued only so long as is necessary to successfully treat the disease or medical condition.

Since compounds of the invention possess AT₁ receptor antagonist activity and/or NEP enzyme inhibition activity, such compounds are also useful as research tools for investigating or studying biological systems or samples having AT₁ receptors or a NEP enzyme, for example to study diseases where the AT₁ receptor or NEP enzyme plays a role. Any suitable biological system or sample having AT₁ receptors and/or a NEP enzyme may be employed in such studies which may be conducted either in vitro or in vivo. Representative biological systems or samples suitable for such studies include, but are not limited to, cells, cellular extracts, plasma membranes, tissue samples, isolated organs, mammals (such as mice, rats, guinea pigs, rabbits, dogs, pigs, humans, and so forth), and the like, with mammals being of particular interest. In one particular embodiment of the invention an AT₁ receptor in a mammal is antagonized by administering an AT₁-antagonizing amount of a compound of the invention. In another particular embodiment, NEP enzyme activity in a mammal is inhibited by administering a NEP-inhibiting amount of a compound of the invention. Compounds of the invention can also be used as research tools by conducting biological assays using such compounds.

When used as a research tool, a biological system or sample comprising an AT₁ receptor and/or a NEP enzyme is typically contacted with an AT₁ receptor-antagonizing or NEP enzyme-inhibiting amount of a compound of the invention. After the biological system or sample is exposed to the compound, the effects of antagonizing the AT₁ receptor and/or inhibiting the NEP enzyme are determined using conventional procedures and equipment, such as by measuring receptor binding in a binding assay or measuring ligand-mediated changes in a functional assay. Exposure encompasses contacting cells or tissue with the compound, administering the compound to a mammal, for example by i.p., i.v. or s.c. administration, and so forth. This determining step can involve measuring a response, i.e., a quantitative analysis or may comprise an observation, i.e., a qualitative analysis. Measuring a response involves, for example, determining the effects of the compound on the biological system or sample using conventional procedures and equipment, such as radioligand binding assays and measuring ligand-mediated changes in functional assays. The assay results can be used to determine the activity level as well as the amount of compound necessary to achieve the desired result, i.e., an AT₁ receptor-antagonizing and/or a NEP enzyme-inhibiting amount. Typically, the determining step will involve determining the AT₁ receptor ligand-mediated effects and/or determining the effects of inhibiting the NEP enzyme.

Additionally, compounds of the invention can be used as research tools for evaluating other chemical compounds, and thus are also useful in screening assays to discover, for example, new compounds having AT₁ receptor-antagonizing activity and/or NEP-inhibiting activity. In this manner, a compound of the invention is used as a standard in an assay to allow comparison of the results obtained with a test compound and with compounds of the invention to identify those test compounds that have about equal or superior activity, if any. For example, K_(i) data (as determined, for example, by a binding assay) for a test compound or a group of test compounds is compared to the Ic data for a compound of the invention to identify those test compounds that have the desired properties, for example, test compounds having a K_(i) value about equal or superior to a compound of the invention, if any. This aspect of the invention includes, as separate embodiments, both the generation of comparison data (using the appropriate assays) and the analysis of test data to identify test compounds of interest. Thus, a test compound can be evaluated in a biological assay, by a method comprising the steps of: (a) conducting a biological assay with a test compound to provide a first assay value; (b) conducting the biological assay with a compound of the invention to provide a second assay value; wherein step (a) is conducted either before, after or concurrently with step (b); and (c) comparing the first assay value from step (a) with the second assay value from step (b). Exemplary biological assays include an AT₁ receptor binding assay and a NEP enzyme inhibition assay.

Pharmaceutical Compositions and Formulations

Compounds of the invention are typically administered to a patient in the form of a pharmaceutical composition or formulation. Such pharmaceutical compositions may be administered to the patient by any acceptable route of administration including, but not limited to, oral, rectal, vaginal, nasal, inhaled, topical (including transdermal), ocular, and parenteral modes of administration. Further, the compounds of the invention may be administered, for example orally, in multiple doses per day (for example, two, three, or four times daily), in a single daily dose or a single weekly dose. It will be understood that any form of the compounds of the invention, (i.e., free base, free acid, pharmaceutically acceptable salt, solvate, etc.) that is suitable for the particular mode of administration can be used in the pharmaceutical compositions discussed herein.

Accordingly, in one embodiment, the invention is directed to a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a compound of the invention. The compositions may contain other therapeutic and/or formulating agents if desired. When discussing compositions, the “compound of the invention” may also be referred to herein as the “active agent,” to distinguish it from other components of the formulation, such as the carrier. Thus, it is understood that the term “active agent” includes compounds of formula I as well as pharmaceutically acceptable salts, solvates and prodrugs of that compound.

The pharmaceutical compositions of the invention typically contain a therapeutically effective amount of a compound of the invention. Those skilled in the art will recognize, however, that a pharmaceutical composition may contain more than a therapeutically effective amount, such as in bulk compositions, or less than a therapeutically effective amount, such as in individual unit doses designed for multiple administration to achieve a therapeutically effective amount. Typically, the composition will contain from about 0.01-95 wt % of active agent, including, from about 0.01-30 wt %, such as from about 0.01-10 wt %, with the actual amount depending upon the formulation itself, the route of administration, the frequency of dosing, and so forth. In one embodiment, a composition suitable for an oral dosage form, for example, may contain about 5-70 wt %, or from about 10-60 wt % of active agent.

Any conventional carrier or excipient may be used in the pharmaceutical compositions of the invention. The choice of a particular carrier or excipient, or combinations of carriers or excipients, will depend on the mode of administration being used to treat a particular patient or type of medical condition or disease state. In this regard, the preparation of a suitable composition for a particular mode of administration is well within the scope of those skilled in the pharmaceutical arts. Additionally, carriers or excipients used in such compositions are commercially available. By way of further illustration, conventional formulation techniques are described in Remington: The Science and Practice of Pharmacy, 20^(th) Edition, Lippincott Williams & White, Baltimore, Md. (2000); and H. C. Ansel et al., Pharmaceutical Dosage Forms and Drug Delivery Systems, 7^(th) Edition, Lippincott Williams & White, Baltimore, Md. (1999).

Representative examples of materials which can serve as pharmaceutically acceptable carriers include, but are not limited to, the following: sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose, such as microcrystalline cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffer solutions; compressed propellant gases, such as chlorofluorocarbons and hydrofluorocarbons; and other non-toxic compatible substances employed in pharmaceutical compositions.

Pharmaceutical compositions are typically prepared by thoroughly and intimately mixing or blending the active agent with a pharmaceutically acceptable carrier and one or more optional ingredients. The resulting uniformly blended mixture may then be shaped or loaded into tablets, capsules, pills, canisters, cartridges, dispensers and the like using conventional procedures and equipment.

In those formulations where the compound of the invention contains a thiol group, additional consideration may be given to minimize or eliminate oxidation of the thiol to form a disulfide. In solid formulations, this may be accomplished by reducing the drying time, decreasing the moisture content of the formulation, and including materials such as ascorbic acid, sodium ascorbate, sodium sulfite and sodium bisulfite, as well as materials such as a mixture of lactose and microcrystalline cellulose. In liquid formulations, stability of the thiol may be improved by the addition of amino acids, antioxidants, or a combination of disodium edetate and ascorbic acid.

In one embodiment, the pharmaceutical compositions are suitable for oral administration. Suitable compositions for oral administration may be in the form of capsules, tablets, pills, lozenges, cachets, dragees, powders, granules; solutions or suspensions in an aqueous or non-aqueous liquid; oil-in-water or water-in-oil liquid emulsions; elixirs or syrups; and the like; each containing a predetermined amount of the active agent.

When intended for oral administration in a solid dosage form (capsules, tablets, pills and the like), the composition will typically comprise the active agent and one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate. Solid dosage forms may also comprise: fillers or extenders, such as starches, microcrystalline cellulose, lactose, sucrose, glucose, mannitol, and/or silicic acid; binders, such as carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and/or acacia; humectants, such as glycerol; disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and/or sodium carbonate; solution retarding agents, such as paraffin; absorption accelerators, such as quaternary ammonium compounds; wetting agents, such as cetyl alcohol and/or glycerol monostearate; absorbents, such as kaolin and/or bentonite clay; lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and/or mixtures thereof; coloring agents; and buffering agents.

Release agents, wetting agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants may also be present in the pharmaceutical compositions. Exemplary coating agents for tablets, capsules, pills and like, include those used for enteric coatings, such as cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropyl methylcellulose phthalate, methacrylic acid-methacrylic acid ester copolymers, cellulose acetate trimellitate, carboxymethyl ethyl cellulose, hydroxypropyl methyl cellulose acetate succinate, and the like. Examples of pharmaceutically acceptable antioxidants include: water-soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfate sodium sulfite and the like; oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, lecithin, propyl gallate, alpha-tocopherol, and the like; and metal-chelating agents, such as citric acid, ethylenediamine tetraacetic acid, sorbitol, tartaric acid, phosphoric acid, and the like.

Compositions may also be formulated to provide slow or controlled release of the active agent using, by way of example, hydroxypropyl methyl cellulose in varying proportions or other polymer matrices, liposomes and/or microspheres. In addition, the pharmaceutical compositions of the invention may contain opacifying agents and may be formulated so that they release the active agent only, or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner. Examples of embedding compositions which can be used include polymeric substances and waxes. The active agent can also be in micro-encapsulated form, if appropriate, with one or more of the above-described excipients.

Suitable liquid dosage forms for oral administration include, by way of illustration, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. Liquid dosage forms typically comprise the active agent and an inert diluent, such as, for example, water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (for example, cottonseed, groundnut, corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Suspensions may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminium metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

When intended for oral administration, the pharmaceutical compositions of the invention may be packaged in a unit dosage form. The term “unit dosage form” refers to a physically discrete unit suitable for dosing a patient, i.e., each unit containing a predetermined quantity of the active agent calculated to produce the desired therapeutic effect either alone or in combination with one or more additional units. For example, such unit dosage forms may be capsules, tablets, pills, and the like.

In another embodiment, the compositions of the invention are suitable for inhaled administration, and will typically be in the form of an aerosol or a powder. Such compositions are generally administered using well-known delivery devices, such as a nebulizer, dry powder, or metered-dose inhaler. Nebulizer devices produce a stream of high velocity air that causes the composition to spray as a mist that is carried into a patient's respiratory tract. An exemplary nebulizer formulation comprises the active agent dissolved in a carrier to form a solution, or micronized and combined with a carrier to form a suspension of micronized particles of respirable size. Dry powder inhalers administer the active agent as a free-flowing powder that is dispersed in a patient's air-stream during inspiration. An exemplary dry powder formulation comprises the active agent dry-blended with an excipient such as lactose, starch, mannitol, dextrose, polylactic acid, polylactide-co-glycolide, and combinations thereof. Metered-dose inhalers discharge a measured amount of the active agent using compressed propellant gas. An exemplary metered-dose formulation comprises a solution or suspension of the active agent in a liquefied propellant, such as a chlorofluorocarbon or hydrofluoroalkane. Optional components of such formulations include co-solvents, such as ethanol or pentane, and surfactants, such as sorbitan trioleate, oleic acid, lecithin, glycerin, and sodium lauryl sulfate. Such compositions are typically prepared by adding chilled or pressurized hydrofluoroalkane to a suitable container containing the active agent, ethanol (if present) and the surfactant (if present). To prepare a suspension, the active agent is micronized and then combined with the propellant. Alternatively, a suspension formulation can be prepared by spray drying a coating of surfactant on micronized particles of the active agent. The formulation is then loaded into an aerosol canister, which forms a portion of the inhaler.

Compounds of the invention can also be administered parenterally (e.g., by subcutaneous, intravenous, intramuscular, or intraperitoneal injection). For such administration, the active agent is provided in a sterile solution, suspension, or emulsion. Exemplary solvents for preparing such formulations include water, saline, low molecular weight alcohols such as propylene glycol, polyethylene glycol, oils, gelatin, fatty acid esters such as ethyl oleate, and the like. Parenteral formulations may also contain one or more anti-oxidants, solubilizers, stabilizers, preservatives, wetting agents, emulsifiers, and dispersing agents. Surfactants, additional stabilizing agents or pH-adjusting agents (acids, bases or buffers) and anti-oxidants are particularly useful to provide stability to the formulation, for example, to minimize or avoid hydrolysis of ester and amide linkages, or dimerization of thiols that may be present in the compound. These formulations may be rendered sterile by use of a sterile injectable medium, a sterilizing agent, filtration, irradiation, or heat. In one particular embodiment, the parenteral formulation comprises an aqueous cyclodextrin solution as the pharmaceutically acceptable carrier. Suitable cyclodextrins include cyclic molecules containing six or more α-D-glucopyranose units linked at the 1,4 positions by a linkages as in amylase, β-cyclodextrin or cycloheptaamylose. Exemplary cyclodextrins include cyclodextrin derivatives such as hydroxypropyl and sulfobutyl ether cyclodextrins such as hydroxypropyl-β-cyclodextrin and sulfobutyl ether β-cyclodextrin. Exemplary buffers for such formulations include carboxylic acid-based buffers such as citrate, lactate and maleate buffer solutions.

Compounds of the invention can also be administered transdermally using known transdermal delivery systems and excipients. For example, the compound can be admixed with permeation enhancers, such as propylene glycol, polyethylene glycol monolaurate, azacycloalkan-2-ones and the like, and incorporated into a patch or similar delivery system. Additional excipients including gelling agents, emulsifiers and buffers, may be used in such transdermal compositions if desired.

If desired, the compounds of the invention may be administered in combination with one or more other therapeutic agents. Thus, in one embodiment, pharmaceutical compositions of the invention contain other drugs that are co-administered with a compound of the invention. For example, the composition may further comprise one or more drugs (also referred to as “secondary agents(s)”) selected from the group of diuretics, β₁ adrenergic receptor blockers, calcium channel blockers, angiotensin-converting enzyme inhibitors, AT₁ receptor antagonists, neprilysin inhibitors, non-steroidal anti-inflammatory agents, prostaglandins, anti-lipid agents, anti-diabetic agents, anti-thrombotic agents, renin inhibitors, endothelin receptor antagonists, endothelin converting enzyme inhibitors, aldosterone antagonists, angiotensin-converting enzyme/neprilysin inhibitors, and combinations thereof. Such therapeutic agents are well known in the art, and examples are described below. By combining a compound of the invention with a secondary agent, triple therapy can be achieved: AT₁ receptor antagonist activity, NEP inhibition activity and activity associated with the secondary agent (for example, β₁ adrenergic receptor blocker) can be achieved using only two active components. Since compositions containing two active components are typically easier to formulate than compositions containing three active components, such two-component compositions provide a significant advantage over compositions containing three active components. Accordingly, in yet another aspect of the invention, a pharmaceutical composition comprises a compound of the invention, a second active agent, and a pharmaceutically acceptable carrier. Third, fourth etc. active agents may also be included in the composition. In combination therapy, the amount of compound of the invention that is administered, as well as the amount of secondary agents, may be less than the amount typically administered in monotherapy.

Compounds of the invention may be either physically mixed with the second active agent to form a composition containing both agents; or each agent may be present in separate and distinct compositions which are administered to the patient simultaneously or at separate times. For example, a compound of the invention can be combined with a second active agent using conventional procedures and equipment to form a combination of active agents comprising a compound of the invention and a second active agent. Additionally, the active agents may be combined with a pharmaceutically acceptable carrier to form a pharmaceutical composition comprising a compound of the invention, a second active agent and a pharmaceutically acceptable carrier. In this embodiment, the components of the composition are typically mixed or blended to create a physical mixture. The physical mixture is then administered in a therapeutically effective amount using any of the routes described herein.

Alternatively, the active agents may remain separate and distinct before administration to the patient. In this embodiment, the agents are not physically mixed together before administration but are administered simultaneously or at separate times as separate compositions. Such compositions can be packaged separately or may be packaged together in a kit. When administered at separate times, the secondary agent will typically be administered less than 24 hours after administration of the compound of the invention, ranging anywhere from concurrent with administration of the compound of the invention to about 24 hours post-dose. This is also referred to as sequential administration. Thus, a compound of the invention can be orally administered simultaneously or sequentially with another active agent using two tablets, with one tablet for each active agent, where sequential may mean being administered immediately after administration of the compound of the invention or at some predetermined time later (for example, one hour later or three hours later). Alternatively, the combination may be administered by different routes of administration, i.e., one orally and the other by inhalation.

In one embodiment, the kit comprises a first dosage form comprising a compound of the invention and at least one additional dosage form comprising one or more of the secondary agents set forth herein, in quantities sufficient to carry out the methods of the invention. The first dosage form and the second (or third, etc,) dosage form together comprise a therapeutically effective amount of active agents for the treatment or prevention of a disease or medical condition in a patient.

Secondary agent(s), when included, are present in a therapeutically effective amount such that they are typically administered in an amount that produces a therapeutically beneficial effect when co-administered with a compound of the invention. The secondary agent can be in the form of a pharmaceutically acceptable salt, solvate, optically pure stereoisomer, and so forth. The secondary agent may also be in the form of a prodrug, for example, a compound having a carboxylic acid group that has been esterified. Thus, secondary agents listed below are intended to include all such forms, and are commercially available or can be prepared using conventional procedures and reagents.

In one embodiment, a compound of the invention is administered in combination with a diuretic. Representative diuretics include, but are not limited to: carbonic anhydrase inhibitors such as acetazolamide and dichlorphenamide; loop diuretics, which include sulfonamide derivatives such as acetazolamide, ambuside, azosernide, bumetanide, butazolamide, chloraminophenamide, clofenamide, clopamide, clorexolone, disulfamide, ethoxolamide, furosemide, mefruside, methazolamide, piretanide, torsemide, tripamide, and xipamide, as well as non-sulfonamide diuretics such as ethacrynic acid and other phenoxyacetic acid compounds such as tienilic acid, indacrinone and quincarbate; osmotic diuretics such as mannitol; potassium-sparing diuretics, which include aldosterone antagonists such as spironolactone, and Na⁺ channel inhibitors such as amiloride and triamterene; thiazide and thiazide-like diuretics such as althiazide, bendroflumethiazide, benzylhydrochlorothiazide, benzthiazide, buthiazide, chlorthalidone, chlorothiazide, cyclopenthiazide, cyclothiazide, epithiazide, ethiazide, fenquizone, flumethiazide, hydrochlorothiazide, hydroflumethiazide, indapamide, methylclothiazide, meticrane, metolazone, paraflutizide, polythiazide, quinethazone, teclothiazide, and trichloromethiazide; and combinations thereof. In a particular embodiment, the diuretic is selected from amiloride, bumetanide, chlorothiazide, chlorthalidone, dichlorphenamide, ethacrynic acid, furosemide, hydrochlorothiazide, hydroflumethiazide, indapamide, methylclothiazide, metolazone, torsemide, triamterene, and combinations thereof. The diuretic will be administered in an amount sufficient to provide from about 5-50 mg per day, more typically 6-25 mg per day, with common dosages being 6.25 mg, 12.5 mg or 25 mg per day.

Compounds of the invention may also be administered in combination with a β₁ adrenergic receptor blocker. Representative β₁ adrenergic receptor blockers include, but are not limited to, acebutolol, alprenolol, amosulalol, arotinolol, atenolol, befunolol, betaxolol, bevantolol, bisoprolol, bopindolol, bucindolol, bucumolol, bufetolol, bufuralol, bunitrolol, bupranolol, bubridine, butofilolol, carazolol, carteolol, carvedilol, celiprolol, cetamolol, cloranolol, dilevalol, epanolol, esmolol, indenolol, labetolol, levobunolol, mepindolol, metipranolol, metoprolol (e.g., metoprolol succinate and metoprolol tartrate), moprolol, nadolol, nadoxolol, nebivalol, nipradilol, oxprenolol, penbutolol, perbutolol, pindolol, practolol, pronethalol, propranolol, sotalol, sufinalol, talindol, tertatolol, tilisolol, timolol, toliprolol, xibenolol, and combinations thereof. In one particular embodiment, the β₁ adrenergic receptor blocker is selected from atenolol, bisoprolol, metoprolol, propranolol, sotalol, and combinations thereof.

In one embodiment, a compound of the invention is administered in combination with a calcium channel blocker. Representative calcium channel blockers include, but are not limited to, amlodipine, anipamil, aranipine, barnidipine, bencyclane, benidipine, bepridil, clentiazem, cilnidipine, cinnarizine, diltiazem, efonidipine, elgodipine, etafenone, felodipine, fendiline, flunarizine, gallopamil, isradipine, lacidipine, lercanidipine, lidoflazine, lomerizine, manidipine, mibefradil, nicardipine, nifedipine, niguldipine, niludipine, nilvadipine, nimodipine, nisoldipine, nitrendipine, nivaldipine, perhexyline, prenylamine, ryosidine, semotiadil, terodiline, tiapamil, verapamil, and combinations thereof. In a particular embodiment, the calcium channel blocker is selected from amlodipine, bepridil, diltiazem, felodipine, isradipine, lacidipine, nicardipine, nifedipine, niguldipine, niludipine, nimodipine, nisoldipine, ryosidine, verapamil, and combinations thereof.

Compounds of the invention can also be administered in combination with an angiotensin-converting enzyme (ACE) inhibitor. Representative ACE inhibitors include, but are not limited to, accupril, alacepril, benazepril, benazeprilat, captopril, ceranapril, cilazapril, delapril, enalapril, enalaprilat, fosinopril, fosinoprilat, imidapril, lisinopril, moexipril, monopril, moveltopril, pentopril, perindopril, quinapril, quinaprilat, ramipril, ramiprilat, saralasin acetate, spirapril, temocapril, trandolapril, zofenopril, and combinations thereof. In a particular embodiment, the ACE inhibitor is selected from: benazepril, enalapril, lisinopril, ramipril, and combinations thereof.

In one embodiment, a compound of the invention is administered in combination with an AT₁ receptor antagonist, also known as angiotensin II type 1 receptor blockers (ARBs). Representative ARBs include, but are not limited to, abitesartan, benzyllosartan, candesartan, candesartan cilexetil, elisartan, embusartan, enoltasosartan, eprosartan, fonsartan, forasartan, glycyllosartan, irbesartan, isoteoline, losartan, medoximil, milfasartan, olmesartan, opomisartan, pratosartan, ripisartan, saprisartan, saralasin, sarmesin, tasosartan, telmisartan, valsartan, zolasartan, and combinations thereof. In a particular embodiment, the ARB is selected from candesartan, eprosartan, irbesartan, losartan, olmesartan, irbesartan, saprisartan, tasosartan, telmisartan, and combinations thereof. Exemplary salts include eprosartan mesylate, losartan potassium salt, and olmesartan medoxomil. Typically, the ARB will be administered in an amount sufficient to provide from about 4-600 mg per dose, with exemplary daily dosages ranging from 20-320 mg per day.

In another embodiment, a compound of the invention is administered in combination with a neprilysin (NEP) inhibitor. Representative NEP inhibitors include, but are not limited to: candoxatril; candoxatrilat; dexecadotril ((+)-N-[2(R)-(acetylthiomethyl)-3-phenylpropionyl]glycine benzyl ester); CGS-24128 (3-[3-(biphenyl-4-yl)-2-(phosphono-methylamino)propionamido]propionic acid); CGS-24592 ((S)-3-[3-(biphenyl-4-yl)-2-(phosphonomethylamino)propionamido]propionic acid); CGS-25155 (N-[9(R)-(acetylthiomethyl)-10-oxo-1-azacyclodecan-2(S)-ylcarbonyl]-4(R)-hydroxy-L-proline benzyl ester); 3-(1-carbamoylcyclohexyl)propionic acid derivatives described in WO 2006/027680 to Hepworth et al. (Pfizer Inc.); JMV-390-1 (2(R)-benzyl-3-(N-hydroxycarbamoyl)-propionyl-L-isoleucyl-L-leucine); ecadotril; phosphoramidon; retrothiorphan; RU-42827 (2-(mercaptomethyl)-N-(4-pyridinyl)benzenepropionamide); RU-44004 (N-(4-morpholinyl)-3-phenyl-2-(sulfanylmethyl)propionamide); SCH-32615 ((S)—N—[N-(1-carboxy-2-phenylethyl)-L-phenylalanyl]-(3-alanine) and its prodrug SCH-34826 ((S)—N—[N-[1-[[(2,2-dimethyl-1,3-dioxolan-4-yl)methoxy]carbonyl]-2-phenylethyl]-L-phenylalanyl]-β-alanine); sialorphin; SCH-42495 (N-[2(S)-(acetylsulfanylmethyl)-3-(2-methylphenyl)-propionyl]-L-methionine ethyl ester); spinorphin; SQ-28132 (N-[2-(mercaptomethyl)-1-oxo-3-phenylpropyl]leucine); SQ-28603 (N-[2-(mercaptomethyl)-1-oxo-3-phenylpropyl]-(3-alanine); SQ-29072 (7-[[2-(mercaptomethyl)-1-oxo-3-phenylpropyl]amino]heptanoic acid); thiorphan and its prodrug racecadotril; UK-69578 (cis-4-[[[1-[2-carboxy-3-(2-methoxyethoxy)propyl]cyclopentyl]carbonyl]amino]cyclohexanecarboxylic acid); UK-447,841 (2-{1-[3-(4-chlorophenyl)propylcarbamoyl]-cyclopentylmethyl}-4-methoxy-butyric acid); UK-505,749 ((R)-2-methyl-3-{1-[3-(2-methylbenzothiazol-6-yl)propylcarbamoyl]cyclopentyl}propionic acid); 5-biphenyl-4-yl-4-(3-carboxypropionylamino)-2-methylpentanoic acid and 5-biphenyl-4-yl-4-(3-carboxypropionylamino)-2-methylpentanoic acid ethyl ester (WO 2007/056546); daglutril [(3S,2′R)-3-{1-[2′-(ethoxy-carbonyl)-4′-phenylbutyl]-cyclopentan-1-carbonylamino}-2,3,4,5-tetrahydro-2-oxo-1H-1-benzazepine-1-acetic acid] described in WO 2007/106708 to Khder et al. (Novartis AG); and combinations thereof. In a particular embodiment, the NEP inhibitor is selected from candoxatril, candoxatrilat, CGS-24128, phosphoramidon, SCH-32615, SCH-34826, SQ-28603, thiorphan, and combinations thereof. The NEP inhibitor will be administered in an amount sufficient to provide from about 20-800 mg per day, with typical daily dosages ranging from 50-700 mg per day, more commonly 100-600 or 100-300 mg per day.

In yet another embodiment, a compound of the invention is administered in combination with a non-steroidal anti-inflammatory agent (NSAID). Representative NSAIDs include, but are not limited to: acemetacin, acetyl salicylic acid, alclofenac, alminoprofen, amfenac, amiprilose, amoxiprin, anirolac, apazone, azapropazone, benorilate, benoxaprofen, bezpiperylon, broperamole, bucloxic acid, carprofen, clidanac, diclofenac, diflunisal, diftalone, enolicam, etodolac, etoricoxib, fenbufen, fenclofenac, fenclozic acid, fenoprofen, fentiazac, feprazone, flufenamic acid, flufenisal, fluprofen, flurbiprofen, furofenac, ibufenac, ibuprofen, indomethacin, indoprofen, isoxepac, isoxicam, ketoprofen, ketorolac, lofemizole, lornoxicam, meclofenamate, meclofenamic acid, mefenamic acid, meloxicam, mesalamine, miroprofen, mofebutazone, nabumetone, naproxen, niflumic acid, oxaprozin, oxpinac, oxyphenbutazone, phenylbutazone, piroxicam, pirprofen, pranoprofen, salsalate, sudoxicam, sulfasalazine, sulindac, suprofen, tenoxicam, tiopinac, tiaprofenic acid, tioxaprofen, tolfenamic acid, tolmetin, triflumidate, zidometacin, zomepirac, and combinations thereof. In a particular embodiment, the NSAID is selected from etodolac, flurbiprofen, ibuprofen, indomethacin, ketoprofen, ketorolac, meloxicam, naproxen, oxaprozin, piroxicam, and combinations thereof.

In yet another embodiment, a compound of the invention is administered in combination with an anti-lipid agent. Representative anti-lipid agents include, but are not limited to, statins such as atorvastatin, fluvastatin, lovastatin, pravastatin, rosuvastatin and simvastatin; cholesteryl ester transfer proteins (CETPs); and combinations thereof.

In yet another embodiment, a compound of the invention is administered in combination with an anti-diabetic agent. Representative anti-diabetic agents include, but are not limited to: injectable drugs such as insulin and insulin derivatives; orally effective drugs including biguanides such as metformin, glucagon antagonists, α-glucosidase inhibitors such as acarbose and miglitol, meglitinides such as repaglinide, oxadiazolidine-diones, sulfonylureas such as chlorpropamide, glimepiride, glipizide, glyburide, and tolazamide, thiazolidinediones such as pioglitazone, and rosiglitazone; and combinations thereof.

In one embodiment, a compound of the invention is administered in combination with an anti-thrombotic agent. Representative anti-thrombotic agents include, but are not limited to, aspirin, anti-platelet agents, heparin, and combinations thereof. Compounds of the invention may also be administered in combination with a renin inhibitor, examples of which include, but are not limited to, aliskiren, enalkiren, remikiren, and combinations thereof. In another embodiment, a compound of the invention is administered in combination with an endothelin receptor antagonist, representative examples of which include, but are not limited to, bosentan, darusentan, tezosentan, and combinations thereof. Compounds of the invention may also be administered in combination with an endothelin converting enzyme inhibitor, examples of which include, but are not limited to, phosphoramidon, CGS 26303, and combinations thereof. In yet another embodiment, a compound of the invention is administered in combination with an aldosterone antagonist, representative examples of which include, but are not limited to, eplerenone, spironolactone, and combinations thereof.

Combined therapeutic agents may also be helpful in further combination therapy with compounds of the invention. For example, a combination of the ACE inhibitor enalapril (in the maleate salt form) and the diuretic hydrochlorothiazide, which is sold under the mark Vaseretic®, or a combination of the calcium channel blocker amlodipine (in the besylate salt form) and the ARB olmesartan (in the medoxomil prodrug form), or a combination of a calcium channel blocker and a statin, all may also be used with the compounds of the invention. Dual-acting agents may also be helpful in combination therapy with compounds of the invention. For example, angiotensin-converting enzyme/neprilysin (ACE/NEP) inhibitors such as: AVE-0848 ((4S,7S,12bR)-7-[3-methyl-2(S)-sulfanylbutyramido]-6-oxo-1,2,3,4,6,7,8,12b-octahydropyrido[2,1-a][2]benzazepine-4-carboxylic acid); AVE-7688 (ilepatril) and its parent compound; BMS-182657 (2-[2-oxo-3(S)-[3-phenyl-2 (S)-sulfanylpropionamido]-2,3,4,5-tetrahydro-1H-1-benzazepin-1-yl]acetic acid); CGS-26303 ([N-[2-(biphenyl-4-yl)-1(S)-(1H-tetrazol-5-yl)ethyl]amino]-methylphosphonic acid); CGS-35601 (N-[1-[4-methyl-2(S)-sulfanylpentanamido]-cyclopentylcarbonyl]-L-tryptophan); fasidotril; fasidotrilate; enalaprilat; ER-32935 ((3R,6S,9aR)-6-[3 (S)-methyl-2 (S)-sulfanylpentanamido]-5-oxoperhydrothiazolo[3,2-a]azepine-3-carboxylic acid); gempatrilat; MDL-101264 ((4S,7S,12bR)-7-[2(S)-(2-morpholinoacetylthio)-3-phenylpropionamido]-6-oxo-1,2,3,4,6,7,8,12b-octahydropyrido[2,1-a][2]benzazepine-4-carboxylic acid); MDL-101287 ([4S-[4a,7a(R*),12bβ]]-7-[2-(carboxymethyl)-3-phenylpropionamido]-6-oxo-1,2,3,4,6,7,8,12b-octahydropyrido[2,1-a][2]benzazepine-4-carboxylic acid); omapatrilat; RB-105 (N-[2(S)-(mercaptomethyl)-3(R)-phenylbutyl]-L-alanine); sampatrilat; SA-898 ((2R,4R)—N-[2-(2-hydroxyphenyl)-3-(3-mercaptopropionyl)thiazolidin-4-ylcarbonyl]-L-phenylalanine); Sch-50690 (N-[1(S)-carboxy-2-[N2-(methanesulfonyl)-L-lysylamino]ethyl]-L-valyl-L-tyrosine); and combinations thereof, may also be included. In one particular embodiment, the ACE/NEP inhibitor is selected from: AVE-7688, enalaprilat, fasidotril, fasidotrilate, omapatrilat, sampatrilat, and combinations thereof.

Other therapeutic agents such as a₂-adrenergic receptor agonists and vasopressin receptor antagonists may also be helpful in combination therapy. Exemplary a₂-adrenergic receptor agonists include clonidine, dexmedetomidine, and guanfacine. Exemplary vasopressin receptor antagonists include tolvaptan.

The following formulations illustrate representative pharmaceutical compositions of the invention.

Exemplary Hard Gelatin Capsules for Oral Administration

A compound of the invention (50g), 440g spray-dried lactose and 10g magnesium stearate are thoroughly blended. The resulting composition is then loaded into hard gelatin capsules (500 mg of composition per capsule). Alternately, a compound of the invention (20 mg) is thoroughly blended with starch (89 mg), microcrystalline cellulose (89 mg) and magnesium stearate (2 mg). The mixture is then passed through a No. 45 mesh U.S. sieve and loaded into a hard gelatin capsule (200 mg of composition per capsule).

Exemplary Gelatin Capsule Formulation for Oral Administration

A compound of the invention (100 mg) is thoroughly blended with polyoxyethylene sorbitan monooleate (50 mg) and starch powder (250 mg). The mixture is then loaded into a gelatin capsule (300 mg of composition per capsule).

Alternately, a compound of the invention (40 mg) is thoroughly blended with microcrystalline cellulose (Avicel PH 103; 260 mg) and magnesium stearate (0.8 mg). The mixture is then loaded into a gelatin capsule (Size #1, White, Opaque) (300 mg of composition per capsule).

Exemplary Tablet Formulation for Oral Administration

A compound of the invention (10 mg), starch (45 mg) and microcrystalline cellulose (35 mg) are passed through a No. 20 mesh U.S. sieve and mixed thoroughly. The granules so produced are dried at 50-60° C. and passed through a No. 16 mesh U.S. sieve. A solution of polyvinylpyrrolidone (4 mg as a 10% solution in sterile water) is mixed with sodium carboxymethyl starch (4.5 mg), magnesium stearate (0.5 mg), and talc (1 mg), and this mixture is then passed through a No. 16 mesh U.S. sieve. The sodium carboxymethyl starch, magnesium stearate and talc are then added to the granules. After mixing, the mixture is compressed on a tablet machine to afford a tablet weighing 100 mg.

Alternately, a compound of the invention (250 mg) is thoroughly blended with microcrystalline cellulose (400 mg), silicon dioxide fumed (10 mg), and stearic acid (5 mg). The mixture is then compressed to form tablets (665 mg of composition per tablet).

Alternately, a compound of the invention (400 mg) is thoroughly blended with cornstarch (50 mg), croscarmellose sodium (25 mg), lactose (120 mg), and magnesium stearate (5 mg). The mixture is then compressed to form a single-scored tablet (600 mg of composition per tablet).

Alternately, a compound of the invention (100 mg) is thoroughly blended with cornstarch (100 mg) with an aqueous solution of gelatin (20 mg). The mixture is dried and ground to a fine powder. Microcrystalline cellulose (50 mg) and magnesium stearate (5 mg) are the admixed with the gelatin formulation, granulated and the resulting mixture compressed to form tablets (100 mg of active per tablet).

Exemplary Suspension Formulation for Oral Administration

The following ingredients are mixed to form a suspension containing 100 mg of active agent per 10 mL of suspension:

Ingredients Amount Compound of the invention 1.0 g Fumaric acid 0.5 g Sodium chloride 2.0 g Methyl paraben 0.15 g Propyl paraben 0.05 g Granulated sugar 25.5 g Sorbitol (70% solution) 12.85 g Veegum ® K (magnesium aluminum silicate) 1.0 g Flavoring 0.035 mL Colorings 0.5 mg Distilled water q.s. to 100 mL

Exemplary Liquid Formulation for Oral Administration

A suitable liquid formulation is one with a carboxylic acid-based buffer such as citrate, lactate and maleate buffer solutions. For example, a compound of the invention (which may be pre-mixed with DMSO) is blended with a 100 mM ammonium citrate buffer and the pH adjusted to pH 5, or with is blended with a 100 mM citric acid solution and the pH adjusted to pH 2. Such solutions may also include a solubilizing excipient such as a cyclodextrin, for example the solution may include 10 wt % hydroxypropyl-β-cyclodextrin.

Other suitable formulations include a 5% NaHCO₃ solution, with or without cyclodextrin.

Exemplary Injectable Formulation for Administration by Injection

A compound of the invention (0.2 g) is blended with 0.4 M sodium acetate buffer solution (2.0 mL). The pH of the resulting solution is adjusted to pH 4 using 0.5 N aqueous hydrochloric acid or 0.5 N aqueous sodium hydroxide, as necessary, and then sufficient water for injection is added to provide a total volume of 20 mL. The mixture is then filtered through a sterile filter (0.22 micron) to provide a sterile solution suitable for administration by injection.

Exemplary Compositions for Administration by Inhalation

A compound of the invention (0.2 mg) is micronized and then blended with lactose (25 mg). This blended mixture is then loaded into a gelatin inhalation cartridge. The contents of the cartridge are administered using a dry powder inhaler, for example.

Alternately, a micronized compound of the invention (10g) is dispersed in a solution prepared by dissolving lecithin (0.2 g) in demineralized water (200 mL). The resulting suspension is spray dried and then micronized to form a micronized composition comprising particles having a mean diameter less than about 1.5 μm. The micronized composition is then loaded into metered-dose inhaler cartridges containing pressurized 1,1,1,2-tetrafluoroethane in an amount sufficient to provide about 10 μg to about 500 μg of the compound of the invention per dose when administered by the inhaler.

Alternately, a compound of the invention (25 mg) is dissolved in citrate buffered (pH 5) isotonic saline (125 mL). The mixture is stirred and sonicated until the compound is dissolved. The pH of the solution is checked and adjusted, if necessary, to pH 5 by slowly adding aqueous 1N sodium hydroxide. The solution is administered using a nebulizer device that provides about 10 μg to about 500 μg of the compound of the invention per dose.

EXAMPLES

The following Preparations and Examples are provided to illustrate specific embodiments of the invention. These specific embodiments, however, are not intended to limit the scope of the invention in any way unless specifically indicated.

The following abbreviations have the following meanings unless otherwise indicated and any other abbreviations used herein and not defined have their standard meaning:

-   -   ACE angiotensin converting enzyme     -   APP aminopeptidase P     -   AT₁ angiotensin II type 1 (receptor)     -   AT₂ angiotensin II type 2 (receptor)     -   BSA bovine serum albumin     -   DCM dichloromethane     -   DIPEA N,N-diisopropylethylamine     -   DMF N,N-dimethylformamide     -   DMSO dimethyl sulfoxide     -   Dnp 2,4-dinitrophenyl     -   DOCA deoxycorticosterone acetate     -   EDC N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride     -   EDTA ethylenediaminetetraacetic acid     -   EGTA ethylene glycol bis((3-aminoethyl         ether)-N,N,N′N′-tetraacetic acid     -   EtOAc ethyl acetate     -   EtOH ethanol     -   HATU N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium         hexafluorophosphate     -   HOBt 1-hydroxybenzotriazole     -   iPrOH isopropyl alcohol     -   Mca (7-methoxycoumarin-4-yl)acyl     -   MeOH methanol     -   NBS N-bromosuccinimide     -   NEP neprilysin (EC 3.4.24.11)     -   PBS phosphate buffered saline     -   SHR spontaneously hypertensive rat     -   TFA trifluoroacetic acid     -   THF tetrahydrofuran     -   Tris tris(hydroxymethyl)aminomethane     -   Tween-20 polyethylene glycol sorbitan monolaurate

Any other abbreviations used herein but not defined have their standard, generally accepted meaning. Unless noted otherwise, all materials, such as reagents, starting materials and solvents, were purchased from commercial suppliers (such as Sigma-Aldrich, Fluka Riedel-de Haen, and the like) and were used without further purification.

Preparation 1 7-Methyl-2-propyl-3H-benzoimidazole-5-carboxylic Acid Methyl Ester

7-Methyl-2-propyl-3H-benzoimidazole-5-carboxylic acid (30.0 g, 137 mmol) was dissolved in MeOH (550 mL) and added to a round bottom flask, followed by the addition of 20 mL of a hydrogen chloride solution (30:70, conc. HCl:water). The mixture was refluxed for 12 hours then concentrated under reduced pressure to afford the title compound as a yellow solid (31.8 g, 137 mmol). MS m/z: [M+H⁺] calcd for C₁₃H₁₆N₂O₂, 233.12. found 233.2.

Preparation 2 4′-Bromomethylbiphenyl-2-carboxylic Acid t-Butyl Ester

A solution of 4′-methylbiphenyl-2-carboxylic acid (48.7 g, 230 mmol) and thionyl chloride (150 mL) was stirred at room temperature. After 5.5 hours, the mixture was concentrated in vacuo. Excess thionyl chloride was removed by co-distillation with toluene to afford a yellow solid (52.6 g). The solid was then dissolved in THF (500 mL) and cooled to 0° C. Potassium t-butoxide (15.0 g, 0.13 mol) was added portion wise, followed by addition of a 1M solution of potassium t-butoxide in THF (250 mL). Additional solid potassium t-butoxide (21.4 g, 100 mmol) was added and the mixture was stirred at 0° C. for 1.5 hours. The mixture was then partitioned between EtOAc and water. The organic layer was washed with saturated aqueous NaCl, dried over MgSO₄, filtered, and concentrated to afford 4′-methylbiphenyl-2-carboxylic acid t-butyl ester (62.3 g) as a yellow oil, which was used directly in the next step.

This oil (62 g, 230 mmol), benzoyl peroxide (3.9 g, 16.0 mmol), and NBS (41.2 g, 230 mmol) were mixed with benzene (800 mL) and heated to reflux. After 4.5 hours, benzoyl peroxide (1g) was added, followed by NBS (16 g, 66.0 mmol) 30 minutes later. The mixture was stirred for a total of 6 hours, then cooled, filtered, and concentrated in vacuo. The resulting residue was crystallized from diethyl ether and hexane at 4° C. overnight to give the title compound (40.7 g) as a pale yellow solid.

¹H NMR (DMSO) δ (ppm) 1.1 (s, 9H), 4.6 (s, 2H), 7.1-7.6 (m, 8H).

Preparation 3 3-(2′-t-Butoxycarbonylbiphenyl-4-ylmethyl)-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic Acid

3-(2′-t-Butoxycarbonylbiphenyl-4-ylmethyl)-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic acid methyl ester (3a): A solution of 7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic acid methyl ester (10.0 g, 43 mmol; prepared as described in Preparation 1) in DMF (300 mL) was cooled in an ice bath. Sodium hydride (4.5 g, 60 wt % in mineral oil) was slowly added over 5 minutes. 4′-Bromomethylbiphenyl-2-carboxylic acid t-butyl ester (15.7 g, 45.2 mmol; purchased from commercial source or prepared as described in Preparation 2) was then added. The mixture was gradually warmed to room temperature, then stirred at 75° C. overnight. The mixture was then cooled and concentrated in vacuo and the resultant residue was partitioned between saturated aqueous NaCl and EtOAc (1:1, 400 mL). The organic layer was collected, dried over MgSO₄ and evaporated to dryness, yielding Intermediate (3a) as a pale yellow oil (16.0 g, 32.1 mmol). MS m/z: [M+H⁺] calcd for C₃₁H₃₄N₂O₄, 499.25. found 499.5.

Intermediate (3a) (9.0 g, 18.1 mmol) and THF (100 mL) were added to a round bottom flask. 1 M NaOH in water (90 mL) was added and the mixture was stirred at room temperature overnight. The mixture was acidified to a pH of 4 with 1M HCl. The solution was then extracted three times with EtOAc, and concentrated under reduced pressure to afford the crude title compound as a yellow solid. MS m/z: [M+H⁺] calcd for C₃₀H₃₂N₂O₄, 485.24. found 485.4.

Preparation 4

((R)-1-Benzyl-2-hydroxycarbamoylethyl)carbamic Acid t-Butyl Ester

DIPEA (32.7 mL, 188 mmol) and EDC (15.4 g, 80.5 mmol) were added to a solution of (R)-3-t-butoxycarbonylamino-4-phenylbutyric acid (15.0 g, 53.7 mmol), HOBt (7.3 g, 53.7 mmol), and hydroxylamine hydrochloride (7.5 g, 107 mmol) in DMF (150 mL). The mixture was stirred at room temperature for 1 day and then concentrated in vacuo to yield a pale yellow oil. The oil was distributed between 5% THF in EtOAc and 1M phosphoric acid. The organic layer was collected and washed with 1M NaOH. The alkaline aqueous layer was then acidified using 1M phosphoric acid and extracted with EtOAc. The organic layer was evaporated and the residue was purified by silica gel chromatography (0-100% EtOAc in hexanes) to afford the title compound as a white solid (1.0 g, 3.4 mmol). MS m/z: [M+H⁺] calcd for C₁₅H₂₂N₂O₄, 295.16. found 295.2.

Example 1 4′-[6-((R)-1-Benzyl-2-hydroxycarbamoylethylcarbamoyl)-4-methyl-2-propylbenzoimidazol-1-ylmethyl]benphinyl-2-carboxylic Acid t-Butyl Ester (1a: R^(1a)=t-butyl) and 4′-[6-((R)-1-Benzyl-2-hydroxycarbamoylethylcarbamoyl)-4-methyl-2-propylbenzoimidazol-1-ylmethyl]biphenyl-2-carboxylic Acid (1b; R^(1a)═H)

To a solution of (R)-1-benzyl-2-hydroxycarbamoylethyl)carbamic acid t-butyl ester (2.8 g, 9.5 mmol; prepared as described in Preparation 4) was added methylene chloride (20 mL) and TFA (30 mL). The mixture was stirred at room temperature for 30 minutes and concentrated to dryness under reduced pressure. The solids were dissolved in DMF (60 mL), followed by the addition of 3-(2′-t-butoxycarbonylbiphenyl-4-ylmethyl)-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic acid (2.6 g, 5.4 mmol; prepared as described in Preparation 3), HOBt (1.3 g, 10 mmol), EDC (1.8 g, 10 mmol;), and DIPEA (3.9 mL, 22.4 mmol). The final mixture was stirred at room temperature overnight, then concentrated under reduced pressure. The residue was dissolved in EtOAc and washed with 1M phosphoric acid , and then a saturated solution of sodium bicarbonate. The organic layer was then concentrated under reduced pressure and purified by reverse phase chromatography (1:1 water:acetonitrile with 0.1% TFA) to obtain compound (1a) as the TFA salt (282 mg, 364 μmol). MS m/z: [M+H⁺] calcd for C₄₀H₄₄N₄O₅, 661.33. found 661.4.

Compound (1a) (282 mg, 364 μmol) was dissolved in methylene chloride (5 mL). TFA (10 mL) was added and the mixture was stirred at room temperature for 2 hours then concentrated in vacuo and purified by reverse phase HPLC (1:1 water/acetonitrile with 0.1% TFA) to obtain compound (1b) as the TFA salt (190 mg, 265 μmol). MS m/z: [M+H⁺] calcd for C₃₆H₃₆N₄O₅, 605.27. found 605.6.

¹H NMR (400 Mz, CD₃OD) 6 (ppm) 1.04 (t, 3H, J=5 Hz), 1.35 (m, 2H), 1.80 (q, 2H, J=5 Hz), 2.43 (m, 2H), 2.67 (s, 3H), 2.94 (d, 2H, J=5 Hz), 3.19 (m, 1H), 5.76 (m, 2H), 7.11 (m, 2H, J=5 Hz), 7.22 (m, 5H), 7.33 (m, 2H), 7.44 (m, 1H), 7.53 (m, 1H), 7.69 s, 1H), 7.81 (m, 1H), 7.87 (s, 1H).

Preparation 5 (R)-2-((R)-2-Amino-3-phenylpropyldisulfanyl)-1-benzylethylamine.2[Hci]

Diisopropyl azodicarboxylate (23.5 mL, 119 mmol) was added to a solution of triphenylphosphine (31.3 g, 119 mmol) in THF (600 mL), cooled in an ice bath. The solution was stirred for 30 minutes, and a white solid was formed. ((R)-1-Benzyl-2-hydroxyethyl)carbamic acid t-butyl (20.0 g, 80 mmol) and thioacetic acid (8.5 mL, 120 mmol) was added. The final mixture was stirred at 0° C. for 1 hour, then stirred at room temperature overnight. The mixture was stripped and diluted with EtOAc (400 mL), and washed with a cold saturated NaHCO₃ solution. The organic layer was dried over MgSO₄, and the filtrate was evaporated to dryness, yielding a pale brown oil. The material was flash chromatographed 30-80% EtOAc:Hex recovering Intermediate (5a) as a brown oil.

Intermediate (5a) was dissolved in 200 mL THF, followed by the addition of 1 M lithium hydroxide in water (398 mL). Air was bubbled vigorously through the solution for 1 hour followed by the addition of 100 mL THF. The air stream was then removed and the mixture was stirred at room temperature for 24 hours. The solution was then acidified to pH 5 with acetic acid, and the THF was removed in vacuo. The precipitate was filtered and rinsed with 50 mL deionized water, producing the dimer (Intermediate 5b). The solid was transferred to a round bottom flask, suspended in 100 mL MeCN, and then concentrated under reduced pressure to remove excess water. The recovered Intermediate 5b was dissolved in 4.0 M of HCl in 1,4-dioxane (150 mL) and stirred at room temperature. After 2 hours, the mixture was concentrated under reduced pressure and the material was triturated with EtOAc. The resulting white solid was filtered, washed with 60 mL EtOAc, and dried in vacuo to yield 8.5 g of the title compound. MS m/z: [M+H⁺] calcd for C₁₈H₂₄N₂S₂ 333.1. found 333.4.

¹H NMR (CD₃OD) δ (ppm): 2.7 (m, 2H), 2.9 (m, 1H), 3.0 (m, 1H), 3.7 (m, 1H), 7.2-7.4 (m, 5H).

Example 2 4′-[6-((R)-1-Benzyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propylbenzoimidazol-1-ylmethyl]biphenyl-2- carboxylic Acid t-Butyl Ester (2a) and 4′-[6-((R)-1-Benzyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propylbenzoimidazol-1-ylmethyl]biphenyl-2-carboxylic Acid (2b)

3-(2′-t-butoxycarbonylbiphenyl-4-ylmethyl)-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic acid (727 mg, 1.5 mmol; prepared as described in Preparation 3) was dissolved into 30 mL DMF. HATU (570 mg, 1.5 mmol) was added and the mixture was stirred for 10 minutes. (R)-2((R)-2-Amino-3-phenylpropyldisulfanyl)-1-benzylethylamine.2[HC1] (304 mg, 0.75 mmol; prepared as described in Preparation 5) was added, followed by the addition of DIPEA (522 μL, 3.0 mmol). The mixture was stirred at room temperature for 5 minutes then warmed to 45° C. After 2 hours, the mixture was cooled to room temperature and partitioned between 100 mL EtOAc and 60 mL 10% LiCl (aqueous). The organic layer was then washed with a saturated NaHCO₃ solution and saturated aqueous NaCl, dried over MgSO₄ and concentrated. The recovered material (1.2 g, 950 mmol; dimer intermediate) was dissolved in THF (30 mL, 0.4 mol). A solution of tris(2-carboxyethyl)phosphine hydrochloride (1.4 g, 4.8 mmol) in water (5.0 mL) was added, and the mixture was stirred at room temperature for 1 hour. The mixture was then quenched with 1 mL acetic acid and diluted with 50 mL EtOAc and 10 mL water. The layers were separated and the organic was washed with saturated aqueous NaCl. EtOAc was removed in vacuo recovering Compound (2a) as an oil, which was immediately dissolved in 30 mL of 30% TFA:DCM and stirred for 2 hours at room temperature. The mixture was concentrated and purified by preparative HPLC to provide Compound (2b) as a white powder (365 mg). MS m/z: [M+H⁺] calcd for C₃₅H₃₅N₃O₃S, 578.24. found 578.5.

¹H NMR (CD₃OD) δ (ppm): 1.0 (t, 3H), 1.8 (m, 2H), 2.6-2.8 (m, 5H), 3.9 (m, 1H), 3.1 (m, 1H), 3.2 (t, 2H), 4.3 (m, 1H), 5.8 (s, 2H), 7.1 (d, 1H), 7.1-7.3 (m, 6H), 7.3-7.4 (m, 3H), 7.4-7.5 (m, 1H), 7.5-7.6 (m, 1H), 7.5-7.6 (m, 1H), 7.7 (s, 1H), 7.8 (d, 1H), 7.9 (s, 1H).

Preparation 6 4-Bromomethyl-2-fluorobenzoic acid Methyl Ester

2-Fluoro-4-methylbenzoic acid methyl ester (6a): 2-Fluoro-4-methylbenzoic acid (30.0 g, 195 mmol) was dissolved in MeOH (600 mL, 10 mol), and 12 M HCl (3.2 mL, 39 mmol) was added. The mixture was heated to reflux and stirred for 48 hours, then cooled and concentrated. The recovered material was dried in vacuo for 24 hours to obtain Intermediate (6a) as a white solid that was used without further purification.

Intermediate (6a) (32 g, 188 mmol), NBS (33.5 g, 188 mmol), and benzoyl peroxide (0.46 g, 1.9 mmol) were dissolved in benzene (550 mL, 6.2 mol) and heated to reflux for 20 hours. The mixture was cooled to room temperature, filtered and concentrated. The residue was flash chromatographed to produce the title compound as a white solid (23.6 g).

¹H NMR (CDCl₃) δ (ppm): 3.9 (s, 3H), 4.4 (s, 2H), 7.1-7.4 (m, 2H), 7.8 (t, 1H).

Preparation 7 (7-Methyl-2-propyl-3H-benzoimidazol-5-yl)methanol

7-Methyl-2-propyl-3H-benzoimidazole-5-carboxylic acid methyl ester (7a): Sulfuric acid (12 mL, 0.22 mol) was added to 7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic acid (30 g, 136 mmol) dissolved in MeOH (670 mL, 16 mol). The mixture was refluxed for 36 hours and concentrated in vacuo. The recovered material was dissolved in 500 ml, EtOAc and washed with a saturated NaHCO₃ solution. The organic layer was dried over MgSO₄ and concentrated to provide Intermediate (7a) as a brown solid (29.1 g). MS m/z: [M+H⁺] calcd for C₁₃H₁₆N₂O₂, 233.1. found 233.2.

1M LiAlH₄ in THF (380 mL, 380 mmol) was added to a round bottom flask that had been oven dried and cooled under nitrogen. The solution was cooled to 0° C. and stirred under nitrogen. A solution of Intermediate (7a) (29.2 g, 125 mmol) in THF (100 mL) was added dropwise via an addition funnel and the mixture was stirred at 0° C. After 1 hour, the mixture was warmed to room temperature and stirred overnight. The mixture was cooled to 0° C. EtOAc (50 mL) was slowly added to quench the remaining LiAlH₄. The reaction was then quenched with 70 mL of saturated NH₄Cl. The mixture was washed with EtOAc (400 mL) and the layers were separated. The organic layer was dried over MgSO₄ and concentrated, recovering a brown solid that was flash chromatographed in 5% MeOH:EtOAc to produce the title compound as a white foam (18.4 g).

¹H NMR (CD₃OD) δ (ppm): 1.1 (t, 3H), 1.9 (m, 2H), 2.6 (s, 3H), 2.9 (t, 2H), 4.7 (s, 2H), 7.0 (s, 1H), 7.4 (s, 1H).

Preparation 8 4-(6-Aminomethyl-4-methyl-2-propylbenzoimidazol-1-ylmethyl)-2-fluorobenzoic Acid Methyl Ester

2-Fluoro-4-(6-hydroxymethyl-4-methyl-2-propyl-benzoimidazol-1-ylmethyl)-benzoic acid methyl ester (8a): (7-Methyl-2-propyl-3H-benzoimidazol-5-yl)methanol (3 g, 15 mmol; prepared as described in Preparation 7) was dissolved in 20 mL of dry DMF. The solution was cooled to 0° C. and sodium hydride (598 mg, 15 mmol, 60 wt % in mineral oil) was added. The mixture was stirred for 30 minutes. 4-Bromomethyl-2-fluorobenzoic acid methyl ester (3.7 g, 15 mmol; prepared as described in Preparation 3) in 50 mL of DMF was added, and the mixture was warmed to room temperature and stirred for 10 minutes. The solution was partitioned between EtOAc (300 mL) and 150 mL 10% LiCl (aqueous). The organic layer was separated, dried over MgSO4 and concentrated. The concentrate was flash chromatographed to produce Intermediate (8a) as a clear oil (2.4 g). MS m/z: [M+H⁺] calcd for C₂₁H₂₃FN₂O₃, 371.7. found 371.0.

4-(6-Azidomethyl-4-methyl-2-propylbenzoimidazol-1-ylmethyl)-2-fluorobenzoic acid methyl ester (8b): Intermediate (8a) (2.4 g, 6.5 mmol) was added to 125 mL of DCM followed by DIPEA (4.6 mL, 26.1 mmol) and 4-dimethylaminopyridine (80 mg, 653 μmol). Methanesulfonyl chloride (1.0 mL, 13.1 mmol) was then added at 0° C. and the mixture was warmed to room temperature. After 3 hours, the mixture was concentrated and the material redissolved in 40 mL DMF. Sodium azide (467 mg, 7.2 mmol) was added and the mixture was heated at 80° C. for 1 hour. The mixture was then cooled to room temperature. The solution was partitioned between EtOAc (100 mL) and 50 mL 10% LiCl (aqueous). The organic layer was concentrated and flash chromatographed 20-70% EtOAc:hexanes to yield Intermediate (8b) as a yellow oil (2.1 g). MS m/z: [M+H⁺] calcd for C₂₁H₂₂FN₅O₂, 396.2. found 396.0.

Intermediate (8b) (1.9 g, 4.9 mmol) was dissolved in THF (24 mL). Triphenylphosphine (1.7 g, 6.3 mmol) was added and the mixture was stirred at room temperature for 30 minutes. Water (175 μL, 9.7 mmol) was added and the mixture was heated to 60° C. After 3 hours, the mixture was cooled to room temperature and concentrated in vacuo. The concentrate was dissolved in EtOAc (50 mL) and washed twice in 30 mL 1N HCl. The aqueous layer was extracted with EtOAc (50 mL) then basified to pH˜10 with 6N NaOH. The solution was saturated with NaCl, extracted four times with 50 mL DCM and dried over MgSO₄. The organic layer was then concentrated to provide the title compound (1.8 g), which was used without further purification.

¹H NMR (CD₃OD) δ (ppm): 0.9 (t, 3H), 1.7 (m, 2H), 2.6 (s, 3H), 2.8 (t, 2H), 4.8 (s, 2H), 4.9 (s, 3H), 5.6 (s, 2H), 6.9 (m, 2H), 7.0 (s, 1H), 7.1 (s, 1H), 7.8 (t, 1H).

Preparation 9 (S)-2-Acetylsulfanyl-4-methylpentanoic Acid

(R)-2-Bromo-4-methylpentanoic acid (9a): D-Leucine (9.9 g, 75.2 mmol) was dissolved in 3M HBr in water (120 mL) and cooled to 0° C. A solution of sodium nitrite (8.3 g, 120 mmol) in water (20 mL, 100 mmol) was added. The mixture was stirred at 0° C. for 2.5 hours, extracted twice with EtOAc, washed twice with saturated aqueous NaCl, dried over MgSO₄, filtered, and concentrated to afford 12.6 g (86%) of Intermediate (9a) as a pale yellow oil. This was taken on to the next step without further purification.

¹H NMR (DMSO) 4.31 (1H, t), 1.75 (2H, m) 1.65 (1H, m) 0.82 (6H, dd).

Intermediate (9a) (12.5 g, 64.1 mmol), potassium thioacetate (11.0 g, 96.1 mmol), and DMF (100 mL, 1.0 mol) were added to a round bottom flask, and the mixture stirred at room temperature for 1 hour. Water (100 mL) was added, and the mixture extracted three times with EtOAc, washed twice with saturated aqueous NaCl, dried over MgSO₄, filtered, and concentrated. The product was purified by silica gel chromatography (0-40% EtOAc: hexanes with 5% acetic acid) to yield the title compound (6.8 g) as a pale yellow oil.

¹H NMR (DMSO) 3.96 (1H, t), 2.45 (3H, s), 1.70 (1H, m), 1.55 (1H, m), 1.42 (1H, m), 0.84 (6H, dd).

Example 3 4-{6-[((S)-2-Acetylsulfanyl-4-methylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}-2-fluorobenzoic Acid Methyl Ester (3a; R^(1a)=—CH₃; R^(5a)=—C(O)CH₃) and 2-Fluoro-4-{6-[((S)-2-mercapto-4-methylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic Acid (3b; R^(1a)═R^(5a)═H)

(S)-2-Acetylsulfanyl-4-methylpentanoic acid (19.0 mg, 0.10 mmol; prepared as described in Preparation 9) was dissolved in DMF (1.0 mL), followed by the addition of EDC (19.1 mg, 100 μmol) and HOBt (13.5 mg, 0.1 mmol). The solution was stirred for 30 minutes. 4-(6-Aminomethyl-4-methyl-2-propylbenzoimidazol-1-ylmethyl)-2-fluorobenzoic acid methyl ester (36.9 mg, 0.1 mmol; prepared as described in Preparation 8) in DMF (0.5 mL) was added, followed by the addition of DIPEA (17.4 μL, 0.1 mmol), and the mixture was stirred for 20 hours to yield Compound (3a). The mixture was concentrated under reduced pressure and then redissolved in MeOH (1 mL). 3N NaOH (750 μL, 23 mmol) was added, under nitrogen, and the mixture was stirred for 3 hours. The mixture was then acidified with 6N HCl (0.5 mL, 30 mmol), concentrated under reduced pressure and then purified by preparative HPLC to yield Compound (3b) as a white solid (0.03 g, 50%). MS m/z: [M+H⁺] calcd for C₂₁H₂₂FN₅O₂, 486.2. found 486.2.

Preparation 10 4′46-Amino-4-methyl-2-propylbenzoimidazol-1-ylmethyl)biphenyl-2-carboxylic Acid

4′46-Azidocarbonyl-4-methyl-2-propylbenzoimidazol-1-ylmethyl)biphenyl-2-carboxylic acid t-butyl ester (10a): DIPEA (0.43 mL, 25 mmol) and diphenylphosphonic azide (489 μL, 2.3 mmol) were added to a solution of 3-(2′-t-butoxycarbonylbiphenyl-4-ylmethyl)-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic acid (1.0 g, 2.1 mmol; prepared as described in Preparation 3) in DMF (15 mL). The mixture was stirred at room temperature for 16 hours, diluted with saturated aqueous NaCl (100 mL), then extracted with EtOAc (200 mL). The organic layer was evaporated to dryness to yield crude Intermediate (10a). MS m/z: [M+H⁺] calcd for C₃₀H₃₁N₅O₃, 510.2. found 510.6.

4′-(6-Amino-4-methyl-2-propylbenzoimidazol-1-ylmethyl)biphenyl-2-carboxylic acid t-butyl ester (10b): Crude intermediate (10a) was suspended in toluene (300 mL), and heated at 80° C. overnight. The reactant was concentrated in vacuo, dissolved in 20% aqueous THF (100 mL), and concentrated again in vacuo to yield Intermediate (10b) as an oil. MS m/z: [M+H⁺] calcd for C₂₉H₃₃N₃O₂, 456.2. found 456.2.

Intermediate (10b) was suspended in concentrated HCl and heated at 100° C. for 3 hours. The solution was concentrated and the residue purified by preparative HPLC to yield the title compound. MS m/z: [M+H⁺] calcd for C₂₅H₂₅N₃O₂, 400.2. found 400.4.

Example 4 4-[6-((S)-2-Acetylsulfanyl-4-methylpentanoylamino)-4-methyl-2-propylbenzoimidazol-1-ylmethyl]biphenyl-2-carboxylic Acid (4a; R^(5a)=—C(O)CH₃) and 4′-[6-((S)-2-Mercapto-4-methylpentanoylamino)-4-methyl-2-propylbenzoimidazol-1-ylmethyl]biphenyl-2-carboxylic Acid (4b; R^(5a)═H)

(S)-2-Acetylsulfanyl-4-methylpentanoic acid (15.2 mg, 0.80 mmol; prepared as described in Preparation 9) was dissolved into DMF (1.0 mL). HATU (30.4 mg, 80 μmol) was added, followed by DIPEA (14 μL, 80 μmol) and the solution was stirred for 15 minutes. 4′-(6-Amino-4-methyl-2-propylbenzoimidazol-1-ylmethyl)biphenyl-2-carboxylic acid (32 mg, 80 μmol; prepared as described in Preparation 10) in DMF (0.5 mL) and DIPEA (28 μL, 160 μmol) was added to this solution and the mixture was shaken at room temperature for 2 hours to give Compound (4a). The mixture was concentrated under reduced pressure and the residue was dissolved in MeOH (1 mL). The mixture was placed under an atmosphere of nitrogen and a solution of 3N NaOH (0.75 mL) was added and the mixture was stirred for 2 hours. The solution was acidified with 0.50 mL 6N HCl and concentrated under reduced pressure. The recovered residue was purified by preparative HPLC to yield Compound (4b) (7.4 mg). MS m/z: [M+H⁺] calcd for C₃₁H₃₅N₃O₃S, 530.24. found 530.2.

Example 5

Following the procedures described in the examples above, and substituting the appropriate starting materials and reagents, compounds 5-1 to 5-31, having the following formula, were also prepared:

Ex. R¹ R⁵ R⁶ 5-1 tetrazol-5-yl —CH₂SH —CH₂CH(CH₃)₂ 5-2 —COOH —SH —(CH₂)₂CH₃ 5-3 —COOH —SH —CH₂CH(CH₃)₂ 5-4 —C(O)OC(CH₃)₃ —SC(O)CH₃ —(CH₂)₂CH₃ 5-5 —C(O)OC(CH₃)₃ —SC(O)CH₃ —CH₂CH(CH₃)₂ 5-6 tetrazol-5-yl —CH₂SH —CH(CH₃)CH₂CH₃ 5-7 tetrazol-5-yl —CH₂SH —CH₂-naphthalen-1-yl 5-8 —COOH —CH₂SH —(CH₂)₃CH₃ 5-9 —COOH —CH₂SH —(CH₂)₂CH(CH₃)₂ 5-10 —COOH —CH₂SH —CH(CH₃)CH₂CH₃ 5-11 —COOH —CH₂SH —CH₂-cyclopentyl 5-12 —COOH —CH₂SH —CH₂-cyclohexyl 5-13 —COOH —CH₂SH —CH₂-naphthalen-1-yl 5-14 —COOH —CH₂SH —(CH₂)₂CH(CH₃)₂ 5-15 —COOH —CH₂SH benzyl 5-16 —COOH —CH₂SH —CH₂-cyclopropyl 5-17 —COOH —CH₂SH —CH(CH₃)₂ 5-18 —COOH —CH₂SH —CH(CH₃)₂ 5-19 —COOH —SH —(CH₂)₂CH₃ 5-20 —COOH —SH —(CH₂)₂CH₃ 5-21 —COOH —SH —CH₂C(CH₃)₃ 5-22 —COOH —CH₂SH —CH₂CH(CH₃)₂ 5-23 —COOH —SH —CH₂C(CH₃)₃ 5-24 —COOH —SH —CH₂CH(CH₃)₂ 5-25 —COOH —SH —CH₂CH(CH₃)₂ 5-26 —COOH —CH₂SH —CH₂CH(CH₃)₂ 5-27 tetrazol-5-yl —SH —CH₂CH(CH₃)₂ 5-28 tetrazol-5-yl —SH —CH₂C(CH₃)₃ 5-29 tetrazol-5-yl —CH₂SH —CH₂CH(CH₃)₂ 5-30 tetrazol-5-yl —SH —CH₂CH(CH₃)₂ 5-31 tetrazol-5-yl —CH₂SH —CH₂CH(CH₃)₂

-   (5-1) 2-mercaptomethyl-4-methylpentanoic acid     {7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]-3H-benzoimidazol-5-ylmethyl}amide.     MS m/z: [M+H⁺] calcd for C₃₃H₃₉N₇OS, 582.29. found 582.4. -   (5-2) 4′-{6-[(2-mercaptop     entanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₁H₃₅N₃O₃S, 530.24. found 530.4. -   (5-3)     4′-{6-[(2-mercapto-4-methylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₂H₃₇N₃O₃S, 544.26. found 544.4. -   (5-4)     4′-{6-[(2-acetylsulfanylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid t-butyl ester. MS m/z: [M+H⁺] calcd for C₃₇H₄₅N₃O₄S, 628.31.     found 628.4. -   (5-5)     4′-{6-[(2-acetylsulfanyl-4-methylpentanoylamino)methyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid t-butyl ester. MS m/z: [M+H⁺]calcd for C₃₈H₄₇N₃O₄S, 642.33.     found 642.4. -   (5-6) 2-mercaptomethyl-3-methylpentanoic acid     {7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]-3H-benzoimidazol-5-ylmethyl}amide.     MS m/z: [M+H⁺] calcd for C₃₃H₃₉N₇OS, 582.29. found 582.4. -   (5-7)     2-mercaptomethyl-N-{7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]-3H-benzoimidazol-5-ylmethyl}-3-naphthalen-1-ylpropionamide.     MS m/z: [M+H⁺] calcd for C₄₀H₃₉N₇OS, 666.29. found 666.4. -   (5-8)     4′-{6-[(2-mercaptomethyl-hexanoylamino)methyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₃H₃₉N₃O₃S, 558.27. found 558.2. -   (5-9)     4′-{6-[(2-mercaptomethyl-5-methyl-hexanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₄H₄₁N₃O₃S, 572.29. found 572.2. -   (5-10)     4′-{6-[(2-mercaptomethyl-3-methylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₃H₃₉N₃O₃S, 558.27. found 558.2. -   (5-11) 4′-{6-[(3-cyclop entyl-2-merc     aptomethyl-propionylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₅H₄₁N₃O₃S, 584.29. found 584.2. -   (5-12)     4′-{6-[(3-cyclohexyl-2-mercaptomethyl-propionylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₆H₄₃N₃O₃S, 598.30. found 598.4. -   (5-13)     4′-{6-[(2-mercaptomethyl-3-naphthalen-1-yl-propionylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₀H₃₉N₃O₃S, 642.27. found 642.2. -   (5-14)     4′-{6-[(2-mercaptomethyl-4-methylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₃H₃₉N₃O₃S, 558.27. found 558.2. -   (5-15)     4′-{6-[(2-mercaptomethyl-3-phenylpropionylamino)methyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₆H₃₇N₃O₃S, 592.26. found 592.2. -   (5-16)     4′-{6-[(3-cyclopropyl-2-mercaptomethyl-propionylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₃H₃₇N₃O₃S, 556.26. found 556.2. -   (5-17)     4′-{6-[((R)-2-mercaptomethyl-3-methyl-butyrylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₂H₃₇N₃O₃S, 544.26. found 544.2. -   (5-18)     4′-{6-[((S)-2-mercaptomethyl-3-methyl-butyrylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₂H₃₇N₃O₃S, 544.26. found 544.2. -   (5-19)     4′-{6-[((S)-2-mercaptopentanoylamino)methyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₁H₃₅N₃O₃S, 530.24. found 530.2. -   (5-20)     4′-{6-[((R)-2-mercaptopentanoylamino)methyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₁H₃₅N₃O₃S, 530.24. found 530.2. -   (5-21)     4′-{6-[((S)-2-mercapto-4,4-dimethylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₃H₃₉N₃O₃S, 558.27. found 558.2. -   (5-22)     4′-{6-[((S)-2-mercaptomethyl-4-methylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₃H₃₉N₃O₃S, 558.27. found 558.2. -   (5-23)     4′-{6-[((R)-2-mercapto-4,4-dimethylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₃H₃₉N₃O₃S, 558.27. found 558.2. -   (5-24)     4′-{6-[((R)-2-mercapto-4-methylpentanoylamino)methyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₂H₃₇N₃O₃S, 544.26. found 544.2. -   (5-25)     4′-{6-[((S)-2-mercapto-4-methylpentanoylamino)methyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₂H₃₇N₃O₃S, 544.26. found 544.2. -   (5-26)     4′-{6-[((R)-2-mercaptomethyl-4-methylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₃H₃₉N₃O₃S, 558.27. found 558.2. -   (5-27) (S)-2-mercapto-4-methylpentanoic acid     {7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]-3H-benzoimidazol-5-ylmethyl}amide.     MS m/z: [M+H⁺] calcd for C₃₂H₃₇N₇OS, 568.28. found 568.2. -   (5-28) (S)-2-mercapto-4,4-dimethylpentanoic acid     {7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]-3H-benzoimidazol-5-ylmethyl}amide.     MS m/z: [M+H⁺] calcd for C₃₃H₃₉N₇OS, 582.29. found 582.2. -   (5-29) (S)-2-mercaptomethyl-4-methylpentanoic acid     {7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]-3H-benzoimidazol-5-ylmethyl}amide.     MS m/z: [M+H⁺] calcd for C₃₃H₃₉N₇OS, 582.29. found 582.2. -   (5-30) (R)-2-mer capto-4-methylpentanoic acid     {7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]-3H-benzoimidazol-5-ylmethyl}amide.     MS m/z: [M+H⁺] calcd for C₃₂H₃₇N₇OS, 568.28. found 568.2. -   (5-31) (R)-2-mercaptomethyl-4-methylpentanoic acid     {7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]-3H-benzoimidazol-5-ylmethyl}amide.     MS m/z: [M+H⁺] calcd for C₃₃H₃₉N₇OS, 582.29. found 582.4.

Example 6

Following the procedures described in the examples above, and substituting the appropriate starting materials and reagents, compounds 6-1 to 6-21, having the following formula, were also prepared:

Ex. R¹ R⁵ R⁶ 6-1 —COOH —SH —CH₂CH(CH₃)₂ 6-2 —COOH —CH₂SH —CH₂CH(CH₃)₂ 6-3 —COOH —CH₂SH benzyl 6-4 —COOH —SH —(CH₂)₂CH₃ 6-5 —COOH —CH₂SH —CH(CH₃)₂ 6-6 —COOH —CH₂SH —CH(CH₃)₂ 6-7 —COOH —CH₂SH —CH₂-cyclopropyl 6-8 —COOH —SH —(CH₂)₂CH₃ 6-9 —COOH —CH₂SH —CH₂CH(CH₃)₂ 6-10 tetrazol-5-yl —CH₂SH benzyl 6-11 tetrazol-5-yl —CH₂SH —(CH₂)₂CH₃ 6-12 tetrazol-5-yl —CH₂SH —CH(CH₃)₂ 6-13 tetrazol-5-yl —SH —CH₂CH(CH₃)₂ 6-14 tetrazol-5-yl —SH benzyl 6-15 tetrazol-5-yl —SH —CH₂-cyclopropyl 6-16 tetrazol-5-yl —CH₂SH —CH₂-cyclopropyl 6-17 tetrazol-5-yl —CH₂SH —CH₂CH(CH₃)₂ 6-18 tetrazol-5-yl —SH —(CH₂)₂CH₃ 6-19 tetrazol-5-yl —CH₂SH —CH(CH₃)₂ 6-20 tetrazol-5-yl —SH —CH₂CH(CH₃)₂ 6-21 tetrazol-5-yl —SH benzyl

-   (6-1)     4′-[6((R)-2-mercapto-4-methylpentanoylamino)-4-methyl-2-propyl-benzoimidazol-1-ylmethylbiphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₁H₃₅N₃O₃S, 530.24. found 530.2. -   (6-2)     4′-[6((S)-2-mercaptomethyl-4-methylpentanoylamino)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₂H₃₇N₃O₃S, 544.26. found 544.2. -   (6-3)     4′-[6((S)-2-benzyl-3-mercaptopropionylamino)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₅H₃₅N₃O₃S, 578.24. found 578.4. -   (6-4) 4′-[6-((S)-2-mercaptop     entanoylamino)-4-methyl-2-propylbenzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₀H₃₃N₃O₃S, 516.22. found 516.4. -   (6-5)     4′-[6((R)-2-mercaptomethyl-3-methylbutyrylamino)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₁H₃₅N₃O₃S, 530.24. found 530.4. -   (6-6)     4′-[6((S)-2-mercaptomethyl-3-methylbutyrylamino)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₁H₃₅N₃O₃S, 530.24. found 530.4. -   (6-7)     4′-[6((R)-2-cyclopropylmethyl-3-mercaptopropionylamino)-4-methyl-2-propylbenzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₂H₃₅N₃O₃S, 542.24. found 542.6. -   (6-8)     4′-[6((R)-2-mercaptopentanoylamino)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₂H₄₁N₃O₃S, 548.29; found 548.2. -   (6-9)     4′-[6((R)-2-mercaptomethyl-4-methylpentanoylamino)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₂H₃₇N₃O₃S, 544.26. found 544.6. -   (6-10)     (S)-2-benzyl-3-mercapto-N-{7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]-3H-benzoimidazol-5-yl}propionamide.     MS m/z: [M+H⁺] calcd for C₃₅H₃₅N₇OS, 602.26. found 602.6. -   (6-11) (S)-2-mer captomethyl-4-methyl-pentanoic acid     {7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]-3H-benzoimidazol-5-yl}amide.     MS m/z: [M+H⁺] calcd for C₃₂H₃₇N₇OS, 568.28. found 568.2. -   (6-12)     (R)-2-mercaptomethyl-3-methyl-N-{7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]-3H-benzoimidazol-5-yl}butyramide.     MS m/z: [M+H⁺] calcd for C₃₁H₃₅N₇O₅, 554.26. found 554.2. -   (6-13) (S)-2-mer capto-4-methylpentanoic acid     {7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]-3H-benzoimidazol-5-yl}amide.     MS m/z: [M+H⁺] calcd for C₃₁H₃₅N₇O₅, 554.26. found 554.2. -   (6-14)     (S)-2-mercapto-N-{7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]-3H-benzoimidazol-5-yl}-3-phenylpropionamide.     MS m/z: [M+H⁺] calcd for C₃₄H₃₃N₇OS, 588.25. found 588.2. -   (6-15)     (S)-3-cyclopropyl-2-mercapto-N-{7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]-3H-benzoimidazol-5-yl}propionamide.     MS m/z: [M+H⁺] calcd for C₃₁H₃₃N₇O₅, 552.25. found 552.2. -   (6-16)     (R)-2-cyclopropylmethyl-3-mercapto-N-{7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]-3H-benzoimidazol-5-yl}propionamide.     MS m/z: [M+H⁺] calcd for C₃₂H₃₅N₇OS, 566.26. found 566.2. -   (6-17) (R)-2-mercaptomethyl-4-methylpentanoic acid     {7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]-3H-benzoimidazol-5-yl}amide.     MS m/z: [M+H⁺] calcd for C₃₂H₃₇N₇OS, 568.28. found 568.2. -   (6-18) (S)-2-mercapto-pentanoic acid     {7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]-3H-benzoimidazol-5-yl}amide.     MS m/z: [M+H⁺] calcd for C₃₀H₃₃N₇OS, 540.25. found 540.2. -   (6-19)     (S)-2-mercaptomethyl-3-methyl-N-{7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]-3H-benzoimidazol-5-yl}butyramide.     MS m/z: [M+H⁺] calcd for C₃₁H₃₅N₇O₅, 554.26. found 554.2. -   (6-20) (R)-2-mercapto-4-methylpentanoic acid     {7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]-3H-benzoimidazol-5-yl}amide.     MS m/z: [M+H⁺] calcd for C₃₁H₃₅N₇O₅, 554.26. found 554.2. -   (6-21)     (R)-2-mercapto-N-{7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]-3H-benzoimidazol-5-yl}-3-phenylpropionamide.     MS m/z: [M+H⁺] calcd for C₃₄H₃₃N₇OS, 588.25. found 588.2.

Example 7

Following the procedures described in the examples above, and substituting the appropriate starting materials and reagents, compounds 7-1 to 7-138, having the following formula, were also prepared:

Ex. R¹ R⁵ R⁶ 7-1 tetrazol-5-yl —CH₂—C(O)N(OH)H benzyl 7-2 tetrazol-5-yl —CH₂SC(O)CH₃ benzyl 7-3 tetrazol-5-yl —CH₂SH benzyl 7-4 —COOH —CH₂SC(O)CH₃ benzyl 7-5 tetrazol-5-yl —CH₂N(OH)C(O)H benzyl 7-6 tetrazol-5-yl —CH₂N(OH)C(O)CH₃ benzyl 7-7 —COOH —CH₂N(OH)C(O)H benzyl 7-8 tetrazol-5-yl —CH₂SH —CH₂CH(CH₃)₂ 7-9 tetrazol-5-yl —CH₂SH —CH₂CH(CH₃)₂ 7-10 —C(O)OC(CH₃)₃ —CH₂C(O)N(OH)H benzyl 7-11 —COOH —CH₂SH —CH₂CH(CH₃)₂ 7-12 —COOH —CH₂N(OH)C(O)H —CH₂CH(CH₃)₂ 7-13 tetrazol-5-yl —CH₂N(OH)C(O)H —CH₂CH(CH₃)₂ 7-14 —C(O)OCH₃ —CH₂SH benzyl 7-15 —COOH —CH₂SC(O)-phenyl benzyl 7-16 —COOH —CH₂SC(O)—CH₂CH₃ benzyl 7-17 —COOH —CH₂—SC(O)—CH₂CH(CH₃)₂ benzyl 7-18 —COOH —CH₂SC(O)—CH(CH₃)₂ benzyl 7-19 —COOH —CH₂—SC(O)—CH₂-cyclopentyl benzyl 7-20 —COOH —CH₂SC(O)-cyclohexyl benzyl 7-21 —COOH —CH₂SC(O)-cyclopentyl benzyl 7-22 —COOH —CH₂SC(O)—C(CH₃)₃ benzyl 7-23 —C(O)OCH₂CH₃ —CH₂SH benzyl 7-24 —C(O)O—(CH₂)₂CH₃ —CH₂SH benzyl 7-25 —C(O)O—CH₂CH(CH₃)₂ —CH₂SH benzyl 7-26 —C(O)O—(CH₂)₃CH₃ —CH₂SH benzyl 7-27 —C(O)O-3-chlorobenzyl —CH₂SH benzyl 7-28 —C(O)O-4-methylbenzyl —CH₂SH benzyl 7-29 —C(O)O-2,6-difluorobenzyl —CH₂SH benzyl 7-30 —C(O)O-3-methoxybenzyl —CH₂SH benzyl 7-31 —C(O)O-benzyl —CH₂SH benzyl 7-32 —C(O)O-3-fluorobenzyl —CH₂SH benzyl 7-33 —C(O)O-4-chlorobenzyl —CH₂SH benzyl 7-34 —C(O)O—CH(CH₃)—OC(O)O-cyclohexyl —CH₂SH benzyl 7-35 —C(O)O—CH(CH₃)—OC(O)O—CH(CH₃)₂ —CH₂SH benzyl 7-36 —COOH —CH₂—SC(O)OCH₂CH₃ benzyl 7-37 —C(O)O—CH(CH₃)—OC(O)O—CH₂CH₃ —CH₂SH benzyl 7-38

—CH₂SH benzyl 7-39 —COOH —CH₂—SC(O)CH(NH₂)CH(CH₃)₂ benzyl 7-40 —COOH —CH₂SC(O)-2-pyrrolidine benzyl 7-41 —COOH —CH₂SH 4-fluorobenzyl 7-42 —COOH —CH₂SH 4-chlorobenzyl 7-43 —COOH —CH₂SH 3-fluorobenzyl 7-44 —COOH —CH₂SH 4-methylbenzyl 7-45 —COOH —CH₂SH 4-bromobenzyl 7-46 —COOH —CH₂SH —CH₂-thiophen-2-yl 7-47 —COOH —CH₂SH —CH₂-cyclohexyl 7-48 —COOH —CH₂SH 2-fluorobenzyl 7-49 —COOH —CH₂SC(O)-2-pyridine benzyl 7-50 —COOH —CH₂SC(O)-4-pyridine benzyl 7-51 —COOH —CH₂SC(O)-3-pyridine benzyl 7-52 —COOH —CH₂SC(O)(CH₂)₂-morpholin-4-yl benzyl 7-53 —COOH —CH₂SC(O)(CH₂)₂-4-methylpiperazin-1-yl benzyl 7-54 —COOH —CH₂SC(O)CH₂-morpholin-4-yl benzyl 7-55 —C(O)O(CH₂)₃- —CH₂SH benzyl morpholin-4-yl 7-56 —COOH —CH₂SH 3-chlorobenzyl 7-57 —COOH —CH₂SH 3,5-difluorobenzyl 7-58 —COOH —CH₂SH 2,4-dichlorobenzyl 7-59 —COOH —CH₂SH 2-chlorobenzyl 7-60 —COOH —CH₂SH 2-trifluoro- methylbenzyl 7-61 —C(O)O—(CH₂)₂CH₃ —CH₂SH —CH₂CH(CH₃)₂ 7-62 —COOH —CH₂SC(O)-2pyridine —CH₂CH(CH₃)₂ 7-63 —COOH —CH₂SC(O)-4-pyridine —CH₂CH(CH₃)₂ 7-64 —COOH —CH₂SC(O)-3-pyridine —CH₂CH(CH₃)₂ 7-65 —COOH —CH₂SC(O)(CH₂)₂-morpholin-4-yl —CH₂CH(CH₃)₂ 7-66 —COOH —CH₂SC(O)(CH₂)₂-4-methylpiperazin-1-yl —CH₂CH(CH₃)₂ 7-67 —COOH —CH₂SC(O)(CH₂)₃-morpholin-4-yl —CH₂CH(CH₃)₂ 7-68 —COOH —CH₂SC(O)CH₂-morpholin-4-yl —CH₂CH(CH₃)₂ 7-69 —COOH —CH₂SC(O)CH₂—OC(O)CH₃ —CH₂CH(CH₃)₂ 7-70 —COOH —CH₂SC(O)CH₃ —CH₂CH(CH₃)₂ 7-71 —COOH —CH₂SC(O)-benzyl —CH₂CH(CH₃)₂ 7-72 —COOH —CH₂SC(O)CH₂CH₃ —CH₂CH(CH₃)₂ 7-73 —COOH —CH₂SC(O)(CH₂)₂CH₃ —CH₂CH(CH₃)₂ 7-74 —COOH —CH₂SC(O)-cyclopentyl —CH₂CH(CH₃)₂ 7-75 —COOH —CH₂SC(O)CH₂-cyclopentyl —CH₂CH(CH₃)₂ 7-76 —COOH —CH₂SC(O)-cyclohexyl —CH₂CH(CH₃)₂ 7-77 —COOH —CH₂SC(O)C(CH₃)₃ —CH₂CH(CH₃)₂ 7-78 —COOH —CH₂SC(O)CH(NH₂)-benzyl benzyl 7-79 —COOH —CH₂SC(O)CH(NH₂)—CH(CH₃)CH₂CH₃ benzyl 7-80 —COOH —CH₂SC(O)CH(NH₂)—CH₂CH(CH₃)₂ benzyl 7-81 —COOH —CH₂SC(O)CH(NH₂)—(CH₂)₂SCH₃ benzyl 7-82 —COOH —CH₂SC(O)CH(NH₂)—CH(OH)(CH₃) benzyl 7-83 —COOH —CH₂SC(O)CH(NH₂)-4-hydroxybenzyl benzyl 7-84 —COOH —CH₂SC(O)CH(NH₂)—C(CH₃)₃ benzyl 7-85 —COOH —CH₂SC(O)CH(NH₂)—CH₂CH₃ benzyl 7-86 —COOH —CH₂SC(O)CH(NH₂)—(CH₂)₂CH₃ benzyl 7-87 —COOH —CH₂SC(O)CH(NH₂)—(CH₂)₃CH₃ benzyl 7-88 —COOH —CH₂SC(O)CH(NH₂)—CH₂-cyclohexyl benzyl 7-89 —COOH —CH₂SC(O)CH(NH₂)—CH₂-cyclobutyl benzyl 7-90 —COOH

benzyl 7-91 —COOH

benzyl 7-92 —COOH

benzyl 7-93 —COOH —CH₂SC(O)-2-aminophenyl benzyl 7-94 —COOH —CH₂SC(O)-3-aminophenyl benzyl 7-95 —COOH —CH₂SC(O)CH(NH₂)-benzyl —CH₂CH(CH₃)₂ 7-96 —COOH —CH₂SC(O)CH(NH₂)—CH(CH₃)CH₂CH₃ —CH₂CH(CH₃)₂ 7-97 —COOH —CH₂SC(O)CH(NH₂)—CH₂CH(CH₃)₂ —CH₂CH(CH₃)₂ 7-98 —COOH —CH₂SC(O)CH(NH₂)—(CH₂)₂SCH₃ —CH₂CH(CH₃)₂ 7-99 —COOH —CH₂SC(O)CH(NH₂)-4-hydroxybenzyl —CH₂CH(CH₃)₂ 7-100 —COOH —CH₂SC(O)CH(NH₂)—C(CH₃)₃ —CH₂CH(CH₃)₂ 7-101 —COOH —CH₂SC(O)CH(NH₂)—CH₂CH₃ —CH₂CH(CH₃)₂ 7-102 —COOH —CH₂SC(O)CH(NH₂)—CH₂-cyclohexyl —CH₂CH(CH₃)₂ 7-103 —COOH —CH₂SC(O)CH(NH₂)—CH₂-cyclobutyl —CH₂CH(CH₃)₂ 7-104 —COOH

—CH₂CH(CH₃)₂ 7-105 —COOH

—CH₂CH(CH₃)₂ 7-106 —COOH

—CH₂CH(CH₃)₂ 7-107 —COOH —CH₂SC(O)-tetrahydrofuran-2-yl —CH₂CH(CH₃)₂ 7-108 —COOH —CH₂SC(O)-2-aminophenyl —CH₂CH(CH₃)₂ 7-109 —COOH —CH₂SC(O)-3-aminophenyl —CH₂CH(CH₃)₂ 7-110 —COOH —CH₂SC(O)-4-aminophenyl —CH₂CH(CH₃)₂ 7-111 —COOH —CH₂SC(O)CH(NH₂)—CH(CH₃)₂ —CH₂CH(CH₃)₂ 7-112 —COOH —CH₂SC(O)-2-pyrrolidine —CH₂CH(CH₃)₂ 7-113 —COOH —CH₂SH 2-fluorobenzyl 7-114 —COOH —CH₂SC(O)CH₂N(CH₃)₂ benzyl 7-115 —COOH —CH₂SC(O)—(CH₂)₂N(CH₃)₂ benzyl 7-116 —COOH —CH₂SC(O)—(CH₂)₃N(CH₃)₂ benzyl 7-117 —COOH —CH₂SC(O)—CH₂-2-methylpiperidin-1-yl benzyl 7-118 —COOH —CH₂SC(O)—CH₂-3-methylpiperidin-1-yl benzyl 7-119 —COOH —CH₂SC(O)—(CH₂)₃N(CH₃)₂ —CH₂CH(CH₃)₂ 7-120 —COOH —CH₂SC(O)—CH₂-2-piperidin-1-yl —CH₂CH(CH₃)₂ 7-121 —COOH —CH₂SC(O)(CH₂)₂COOH benzyl 7-122 —COOH —CH₂SC(O)-4-carboxyphenyl benzyl 7-123 —COOH —CH₂SC(O)—CH(NH₂)(CH₂)₂COOH benzyl 7-124 —COOH —CH₂SC(O)—CH₂CH(NH₂)COOH benzyl 7-125 —COOH —CH₂SC(O)-3-carboxyphenyl benzyl 7-126 —COOH —CH₂SC(O)(CH₂)₂COOH —CH₂CH(CH₃)₂ 7-127 —COOH —CH₂SC(O)-3-carboxyphenyl —CH₂CH(CH₃)₂ 7-128 —COOH —CH₂SC(O)-4-carboxyphenyl —CH₂CH(CH₃)₂ 7-129 —COOH —CH₂SC(O)CH(NH₂)—(CH₂)₂COOH —CH₂CH(CH₃)₂ 7-130 —COOH —CH₂SC(O)—CH₂CH(NH₂)—COOH —CH₂CH(CH₃)₂ 7-131 —COOH —CH₂—N(OH)C(O)H 3-fluorobenzyl 7-132 —COOH —CH₂—N(OH)C(O)H 2-fluorobenzyl 7-133 —COOH —CH₂—N(OH)C(O)H 4-methylbenzyl 7-134 —COOH —CH₂SC(O)—CH(NH₂)—CH₂COOH benzyl 7-135 —COOH —CH₂SC(O)—(CH₂)₂CH(NH₂)—COOH benzyl 7-136 —COOH —CH₂SC(O)—(CH₂)₂CH(NH₂)—COOH —CH₂CH(CH₃)₂ 7-137 —COOH —CH₂SC(O)-CH(NH₂)—CH₂COOH —CH₂CH(CH₃)₂ 7-138 —COOH —CH₂SC(O)CH₂N(CH₃)₂ —CH₂CH(CH₃)₂

-   (7-1)     7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]-3H-benzoimidazole-5-carboxylic     acid ((R)-1-benzyl-2-hydroxycarbamoylethyl)amide. MS m/z: [M+H⁺]     calcd for C₃₆H₃₆N₈O₃, 629.29. found 629.6. -   (7-2) thioacetic acid     S—[(R)-2-({7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]-3H-benzoimidazole-5-carbonyl]amino)-3-phenylpropyl}ester.     MS m/z: [M+H⁺] calcd for C₃₇H₃₇N₇O₂S, 644.27. found 644.3. -   (7-3)     7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]-3H-benzoimidazole-5-carboxylic     acid ((R)-1-benzyl-2-mercaptoethyl)amide. MS m/z: [M+H⁺] calcd for     C₃₅H₃₅N₇OS, 602.26. found 602.6. -   (7-4)     4′-[6-((R)-2-acetylsulfanyl-1-benzylethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₇H₃₇N₃O₄S, 620.25. found 620.8. -   (7-5)     7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]-3H-benzoimidazole-5-carboxylic     acid [(R)-1-benzyl-2-(formylhydroxyamino)ethyl]amide. MS m/z: [M+H⁺]     calcd for C₃₆H₃₆N₈O₃, 629.29. found 629.6. -   (7-6)     7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]-3H-benzoimidazole-5-carboxylic     acid [(R)-2-(acetylhydroxyamino)-1-benzylethyl]amide. MS m/z: [M+H⁺]     calcd for C₃₇H₃₈N₈O₃, 643.31. found 643.5. -   (7-7)     4′-{6-[(R)-1-benzyl-2-(formylhydroxyamino)ethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₆H₃₆N₄O₅, 605.27. found 605.6. -   (7-8)     7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]-3H-benzoimidazole-5-carboxylic     acid ((R)-1-mercaptomethyl-3-methyl-butyl)-amide. MS m/z: [M+H⁺]     calcd for C₃₂H₃₇N₇OS, 568.28. found 568.6. -   (7-9)     7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]-3H-benzoimidazole-5-carboxylic     acid ((S)-1-mercaptomethyl-3-methylbutyl)amide. MS m/z: [M+H⁺] calcd     for C₃₂H₃₇N₇OS, 568.28. found 568.6. -   (7-10)     4′-[6-((R)-1-benzyl-2-hydroxycarbamoyl-ethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid t-butyl ester. MS m/z: [M+H⁺] calcd for C₄₀H₄₄N₄O₅, 661.33.     found 661.4. -   (7-11)     4′-[6-((R)-1-mercaptomethyl-3-methyl-butylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₂H₃₇N₃O₃S, 544.26. found 544.4. -   (7-12)     {(R)-1-[(formylhydroxyamino)methyl]-3-methyl-butylcarbamoyl}-4-methyl-2-propylbenzoimidazol-1-ylmethyl)biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₃H₃₈N₄O₅, 571.28. found 571.6. -   (7-13)     7-methyl-2-propyl-3-[2′-(1H-tetrazol-5-yl)biphenyl-4-ylmethyl]-3H-benzoimidazole-5-carboxylic     acid {(R)-1-[(formylhydroxyamino)methyl]-3-methylbutyl}amide. MS     m/z: [M+H⁺] calcd for C₃₃H₃₈N₈O₃, 595.31. found 595.5. -   (7-14)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid methyl ester. MS m/z: [M+H⁺] calcd for C₃₆H₃₇N₃O₃S, 592.26.     found 592.8. -   (7-15)     4′-[6-((R)-2-benzoylsulfanyl-1-benzylethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₂H₃₉N₃O₄S, 682.27. found 682.2. -   (7-16)     4′-[6-((R)-1-benzyl-2-propionylsulfanyl-ethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₈H₃₉N₃O₄S, 634.27. found 634.2. -   (7-17)     4′-{6-[(R)-1-benzyl-2-(3-methyl-butyrylsulfanyl)ethylcarbamoyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₀H₄₃N₃O₄S, 662.30. found 662.2. -   (7-18)     4′-[6-((R)-1-benzyl-2-isobutyrylsulfanylethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₉H₄₁N₃O₄S, 648.28. found 648.2. -   (7-19)     4′-{6-[(R)-1-benzyl-2-(2-cyclopentylacetylsulfanyl)ethylcarbamoyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₂H₄₅N₃O₄S, 688.31. found 688.2. -   (7-20)     4′-[6-((R)-1-benzyl-2-cyclohexanecarbonylsulfanylethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₂H₄₅N₃O₄S, 688.31. found 688.2. -   (7-21)     4′-[6-((R)-1-benzyl-2-cyclopentanecarbonylsulfanylethyl-carbamoyl)-4-methyl-2-propylbenzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₁H₄₃N₃O₄S, 674.30. found 674.2. -   (7-22)     4′-{6-[(R)-1-benzyl-2-(2,2-dimethylpropionylsulfanyl)ethyl-carbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₀H₄₃N₃O₄S, 662.30. found 662.2. -   (7-23) 4′-[6-((R)-1-Benzyl-2-mercapto     ethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid ethyl ester. MS m/z: [M+H⁺] calcd for C₃₇H₃₉N₃O₃S, 606.27.     found 606.6. -   (7-24) 4′-[6-((R)-1-Benzyl-2-mercapto     ethylcarbamoyl)-4-methyl-2-propylbenzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid propyl ester. MS m/z: [M+H⁺] calcd for C₃₈H₄₁N₃O₃S, 620.29.     found 620.2. -   (7-25) 4′-[6-((R)-1-Benzyl-2-mercapto     ethylcarbamoyl)-4-methyl-2-propylbenzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid isobutyl ester. MS m/z: [M+H⁺] calcd for C₃₉H₄₃N₃O₃S, 634.30.     found 634.2. -   (7-26)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid butyl ester. MS m/z: [M+H⁺] calcd for C₃₉H₄₃N₃O₃S, 634.30.     found 634.2. -   (7-27)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid 3-chlorobenzyl ester. MS m/z: [M+H⁺] calcd for C₄₂H₄₀ClN₃O₃S,     702.25. found 702.2. -   (7-28)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid 4-methylbenzyl ester. MS m/z: [M+H⁺] calcd for C₄₃H₄₃N₃O₃S,     682.30. found 682.2. -   (7-29)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid 2,6-difluorobenzyl ester. MS m/z: [M+H⁺] calcd for     C₄₂H₃₉F₂N₃O₃S, 704.27. found 704.2. -   (7-30)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid 3-methoxybenzyl ester. MS m/z: [M+H⁺] calcd for C₄₃H₄₃N₃O₄S,     698.30. found 698.2. -   (7-31) 4′-[6-((R)-1-benzyl-2-mercapto     ethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid benzyl ester. MS m/z: [M+H⁺] calcd for C₄₂H₄₁N₃O₃S, 668.29.     found 668.2. -   (7-32)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid 3-fluorobenzyl ester. MS m/z: [M+H⁺] calcd for C₄₂H₄₀FN₃O₃S,     686.28. found 686.2. -   (7-33)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid 4-chlorobenzyl ester. MS m/z: [M+H⁺] calcd for C₄₂H₄₀ClN₃O₃S,     702.25. found 702.4. -   (7-34)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid 1-cyclohexyloxycarbonyloxyethyl ester. MS m/z: [M+H⁺] calcd for     C₄₄H₄₉N₃O₆S, 748.33. found 748.0. -   (7-35)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid 1-isopropoxycarbonyloxyethyl ester. MS m/z: [M+H⁺] calcd for     C₄₁H₄₅N₃O₆S, 708.30. found 708.4. -   (7-36)     4′-[6-((R)-1-benzyl-2-ethoxycarbonylsulfanylethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₈H₃₉N₃O₅S, 650.26. found 650.2. -   (7-37)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid 1-ethoxycarbonyloxyethyl ester. MS m/z: [M+H⁺] calcd for     C₄₀H₄₃N₃O₆S, 694.29. found 694.0. -   (7-38)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid 3-oxo-1,3-dihydroisobenzofuran-1-yl ester. MS m/z: [M+H⁺] calcd     for C₄₃H₃₉N₃O₅S, 710.26. found 710.8. -   (7-39)     4′-{6-[(R)-2-((S)-2-amino-3-methylbutyrylsulfanyl)-1-benzylethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₀H₄₄N₄O₄S, 677.31. found 677.4. -   (7-40)     4′-{6-[(R)-1-benzyl-2-((S)-pyrrolidine-2-carbonylsulfanyl)ethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₀H₄₂N₄O₄S, 675.29. found 675.6. -   (7-41)     4′-{6-[(R)-2-(4-fluorophenyl)-1-mercaptomethylethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₅H₃₄FN₃O₃S, 596.23. found 596.4. -   (7-42)     4′-{6-[(R)-2-(4-chlorophenyl)-1-mercaptomethylethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₅H₃₄ClN₃O₃S, 612.20. found 612.3. -   (7-43)     4′-{6-[(R)-2-(3-fluorophenyl)-1-mercaptomethylethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₅H₃₄FN₃O₃S, 596.23. found 596.4. -   (7-44)     4′-[6-((R)-1-mercaptomethyl-2-p-tolylethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₆H₃₇N₃O₃S, 592.26. found 592.5. -   (7-45)     4′-{6-[(R)-2-(4-bromophenyl)-1-mercaptomethylethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₅H₃₄BrN₃O₃S, 656.15. found 658.3     (+2 isotopic effect due to bromide). -   (7-46)     4′-[6-((R)-2-mercapto-1-thiophen-2-ylmethylethylcarbamoyl)-4-methyl-2-propylbenzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₃H₃₃N₃O₃S₂, 584.20. found 584.0. -   (7-47)     4′-[6-((R)-1-cyclohexylmethyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₅H₄₁N₃O₃S, 584.29. found 584.4. -   (7-48)     4′-{6-[(R)-1-(2-fluorobenzyl)-2-mercaptoethylcarbamoyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₅H₃₄FN₃O₃S, 596.23. found 596.6. -   (7-49)     4′-{6-[(R)-1-benzyl-2-(pyridine-2-carbonylsulfanyl)ethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₁H₃₈N₄O₄S, 683.26. found 683.2. -   (7-50)     4′-{6-[(R)-1-benzyl-2-(pyridine-4-carbonylsulfanyl)ethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₁H₃₈N₄O₄S, 683.26. found 683.2. -   (7-51)     4′-{6-[(R)-1-benzyl-2-(pyridine-3-carbonylsulfanyl)ethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₁H₃₈N₄O₄S, 683.26. found 683.2. -   (7-52)     4′-{6-[(R)-1-benzyl-2-(3-morpholin-4-ylpropionyl-sulfanyl)ethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₂H₄₆N₄O₅S, 719.32. found 719.2. -   (7-53)     4′-(6-{(R)-1-benzyl-2-[3-(4-methylpiperazin-1-yl)propionyl-sulfanyl]ethylcarbamoyl}-4-methyl-2-propylbenzoimidazol-1-ylmethyl)biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₃H₄₉N₅O₄S, 732.35. found 732.4. -   (7-54)     4′-{6-[(R)-1-benzyl-2-(2-morpholin-4-ylacetylsulfanyl)ethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₁H₄₄N₄O₅S, 705.30. found 705.2. -   (7-55)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid 3-morpholin-4-yl-propyl ester. MS m/z: [M+H⁺] calcd for     C₄₂H₄₈N₄O₄S, 705.34. found 705.8. -   (7-56)     4′-{6-[(R)-1-(3-chlorobenzyl)-2-mercaptoethylcarbamoyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₅H₃₄ClN₃O₃S, 612.20. found 612.4. -   (7-57)     4′-{6-[(R)-1-(3,5-difluorobenzyl)-2-mercaptoethylcarbamoyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₅H₃₃F₂N₃O₃S, 614.22. found 614.4. -   (7-58)     4′-{6-[(R)-1-(2,4-dichlorobenzyl)-2-mercaptoethylcarbamoyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₅H₃₃Cl₂N₃O₃S, 646.16. found 646.4. -   (7-59)     4′-{6-[(R)-1-(2-chlorobenzyl)-2-mercaptoethylcarbamoyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₅H₃₄ClN₃O₃S, 612.20. found 612.2. -   (7-60)     4′-{6-[(R)-2-mercapto-1-(2-trifluoromethylbenzyl)ethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₆H₃₄F₃N₃O₃S, 646.23. found 646.6. -   (7-61)     4′-[6-((R)-1-mercaptomethyl-3-methylbutylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid propyl ester. MS m/z: [M+H⁺] calcd for C₃₅H₄₃N₃O₃S, 586.30.     found 586.2. -   (7-62)     4′-{4-methyl-6-[(R)-3-methyl-1-(pyridine-2-carbonylsulfanylmethyl)butylcarbamoyl]-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₈H₄₀N₄O₄S, 649.28. found 649.2. -   (7-63)     4′-{4-methyl-6-[(R)-3-methyl-1-(pyridine-4-carbonylsulfanylmethyl)butylcarbamoyl]-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₈H₄₀N₄O₄S, 649.28. found 649.2. -   (7-64)     4′-{4-methyl-6-[(R)-3-methyl-1-(pyridine-3-carbonylsulfanylmethyl)butylcarbamoyl]-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₈H₄₀N₄O₄S, 649.28. found 649.2. -   (7-65) 4′-{4-methyl-6-[(R)-3-methyl-1-(3-morp     holin-4-yl-propionylsulfanylmethyl)-butylcarbamoyl]-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₉H₄₈N₄O₅S, 685.33. found 685.4. -   (7-66)     4′-(4-methyl-6-{(R)-3-methyl-1-[3-(4-methylpiperazin-1-yl)-propionylsulfanylmethyl]-butylcarbamoyl}-2-propylbenzoimidazol-1-ylmethyl)biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₀H₅₁N₅O₄S, 698.37. found 698.4. -   (7-67)     4′-{4-methyl-6-[(R)-3-methyl-1-(4-morpholin-4-yl-butyrylsulfanylmethyl)-butylcarbamoyl]-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₀H₅₀N₄O₅S, 699.35. found 699.4. -   (7-68)     4′-{4-methyl-6-[(R)-3-methyl-1-(2-morpholin-4-yl-acetylsulfanylmethyl)-butylcarbamoyl]-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₈H₄₆N₄O₅S , 671.32. found 671.2. -   (7-69)     4′-{6-[(R)-1-(2-acetoxy-acetylsulfanylmethyl)-3-methyl-butylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₆H₄₁N₃O₆S, 644.27. found 644.2. -   (7-70)     4′-[6-((R)-1-acetylsulfanylmethyl-3-methylbutylcarbamoyl)-4-methyl-2-propylbenzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₄H₃₉N₃O₄S, 586.27. found 586.2. -   (7-71)     4′-[6-((R)-1-benzoylsulfanylmethyl-3-methylbutylcarbamoyl)-4-methyl-2-propylbenzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₉H₄₁N₃O₄S, 648.28. found 648.2. -   (7-72)     4′-[4-methyl-6-((R)-3-methyl-1-propionylsulfanylmethylbutylcarbamoyl)-2-propylbenzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₅H₄₁N₃O₄S, 600.28. found 600.2. -   (7-73)     4′-[6-((R)-1-butyrylsulfanylmethyl-3-methylbutylcarbamoyl)-4-methyl-2-propylbenzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₆H₄₃N₃O₄S, 614.30. found 614.2. -   (7-74) 4′-[6-((R)-1-cyclop     entanecarbonylsulfanylmethyl-3-methylbutylcarbamoyl)-4-methyl-2-propylbenzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₈H₄₅N₃O₄S, 640.31. found 640.2. -   (7-75) 4′-{6-[(R)-1-(2-cyc lop     entylacetylsulfanylmethyl)-3-methylbutylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₉H₄₇N₃O₄S, 654.33. found 654.4. -   (7-76) 4′-[6-((R)-1-cyclohexane     carbonylsulfanylmethyl-3-methyl-butylcarbamoyl)-4-methyl-2-propylbenzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₉H₄₇N₃O₄S, 654.33. found 654.2. -   (7-77)     4′-{6-[(R)-1-(2,2-dimethyl-propionylsulfanylmethyl)-3-methylbutylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₇H₄₅N₃O₄S, 628.31. found 628.2. -   (7-78)     4′-{6-[(R)-2-((S)-2-amino-3-phenyl-propionylsulfanyl)-1-benzylethyl-carbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₄H₄₄N₄O₄S, 725.31. found 725.2. -   (7-79)     4′-{6-[(R)-2-((S)-2-amino-3-methyl-pentanoylsulfanyl)-1-benzylethyl-carbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₁H₄₆N₄O₄S, 691.32. found 691.4. -   (7-80)     4′-{6-[(R)-2-((S)-2-amino-4-methylpentanoylsulfanyl)-1-benzylethyl-carbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₁H₄₆N₄O₄S, 691.32. found 691.4. -   (7-81)     4′-{6-[(R)-2-((S)-2-amino-4-methylsulfanylbutyrylsulfanyl)-1-benzylethyl-carbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₀H₄₄N₄O₄S₂, 709.28. found 709.2. -   (7-82)     4′-{6-[(R)-2-((S)-2-amino-3-hydroxybutyrylsulfanyl)-1-benzylethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₉H₄₂N₄O₅S, 679.29. found 679.2. -   (7-83)     4′-(6-{(R)-2-[(S)-2-amino-3-(4-hydroxyphenyl)propionylsulfanyl]-1-benzylethylcarbamoyl}-4-methyl-2-propylbenzoimidazol-1-ylmethyl)biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₄H₄₄N₄O₅S, 741.30. found 741.2. -   (7-84)     4′-{6-[(R)-2-((S)-2-amino-3,3-dimethylbutyrylsulfanyl)-1-benzylethyl-carbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₁H₄₆N₄O₄S, 691.32. found 691.4. -   (7-85)     4′-{6-[(R)-2-((S)-2-aminobutyrylsulfanyl)-1-benzylethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₉H₄₂N₄O₄S, 663.29. found 663.2. -   (7-86)     4′-{6-[(R)-2-((S)-2-aminopentanoylsulfanyl)-1-benzylethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₀H₄₄N₄O₄S, 677.31. found 677.4. -   (7-87)     4′-{6-[(R)-2-((S)-2-aminohexanoylsulfanyl)-1-benzylethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₁H₄₆N₄O₄S, 691.32. found 691.4. -   (7-88)     4′-{6-[(R)-2-((S)-2-amino-3-cyclohexylpropionylsulfanyl)-1-benzylethyl-carbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₄H₅₀N₄O₄S, 731.36. found 731.4. -   (7-89)     4′-{6-[(R)-2-((S)-2-amino-3-cyclobutylpropionylsulfanyl)-1-benzylethyl-carbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₂H₄₆N₄O₄S, 703.32. found 703.4. -   (7-90)     4′-{6-[(R)-2-(1-aminocyclobutanecarbonylsulfanyl)-1-benzylethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₀H₄₂N₄O₄S, 675.29. found 675.2. -   (7-91)     4′-{6-[(R)-2-(1-aminocyclopentanecarbonylsulfanyl)-1-benzylethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₁H₄₄N₄O₄S, 689.31. found 689.2. -   (7-92)     4′-{6-[(R)-2-(1-aminocyclohexanecarbonylsulfanyl)-1-benzylethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₂H₄₆N₄O₄S, 703.32. found 703.2. -   (7-93)     4′-{6-[(R)-2-(2-aminobenzoylsulfanyl)-1-benzylethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₂H₄₀N₄O₄S, 697.28. found 697.2. -   (7-94)     4′-{6-[(R)-2-(3-aminobenzoylsulfanyl)-1-benzylethylcarbamoyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₂H₄₀N₄O₄S, 697.28. found 697.2. -   (7-95)     4′-{6-[(R)-1-((S)-2-amino-3-phenylpropionylsulfanylmethyl)-3-methylbutyl-carbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₁H₄₆N₄O₄S, 691.32. found 691.4. -   (7-96)     4′-{6-[(R)-1-((S)-2-amino-3-methylpentanoylsulfanylmethyl)-3-methylbutyl-carbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₈H₄₈N₄O₄S, 657.34. found 657.4. -   (7-97)     4′-{6-[(R)-1-((S)-2-amino-4-methylpentanoylsulfanylmethyl)-3-methylbutyl-carbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₈H₄₈N₄O₄S, 657.34. found 657.4. -   (7-98)     4′-{6-[(R)-1-((S)-2-amino-4-methylsulfanylbutyrylsulfanylmethyl)-3-methylbutylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₇H₄₆N₄O₄S₂, 675.30. found 675.2. -   (7-99)     4′-(6-{(R)-1-[(S)-2-amino-3-(4-hydroxyphenyl)-propionylsulfanylmethyl]-3-methyl-butylcarbamoyl}-4-methyl-2-propylbenzoimidazol-1-ylmethyl)biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₁H₄₆N₄O₅S, 707.32. found 707.2. -   (7-100)     4′-{6-[(R)-1-((S)-2-amino-3,3-dimethyl-butyrylsulfanylmethyl)-3-methylbutylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₈H₄₈N₄O₄S, 657.34. found 657.4. -   (7-101)     4′-{6-[(R)-1-((S)-2-aminobutyrylsulfanylmethyl)-3-methyl-butylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₆H₄₄N₄O₄S, 629.31. found 629.2. -   (7-102)     4′-{6-[(R)-1-((S)-2-amino-3-cyclohexylpropionylsulfanylmethyl)-3-methylbutylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₁H₅₂N₄O₄S, 697.37. found 697.4. -   (7-103)     4′-{6-[(R)-1-((S)-2-amino-3-cyclobutyl-propionylsulfanylmethyl)-3-methylbutylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₉H₄₈N₄O₄S, 669.34. found 669.4. -   (7-104)     4′-{6-[(R)-1-(1-aminocyclobutanecarbonylsulfanylmethyl)-3-methylbutyl-carbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₇H₄₄N₄O₄S, 641.31. found 641.2. -   (7-105)     4′-{6-[(R)-1-(1-aminocyclopentanecarbonylsulfanylmethyl)-3-methylbutyl-carbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₈H₄₆N₄O₄S, 655.32. found 655.4. -   (7-106)     4′-{6-[(R)-1-(1-aminocyclohexanecarbonylsulfanylmethyl)-3-methylbutyl-carbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₉H₄₈N₄O₄S, 669.34. found 669.4. -   (7-107)     4′-{4-methyl-6-[(R)-3-methyl-1-((S)-tetrahydrofuran-2-carbonylsulfanyl-methyl)-butylcarbamoyl]-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₇H₄₃N₃O₅S, 642.29. found 642.2. -   (7-108)     4′-{6-[(R)-1-(2-aminobenzoylsulfanylmethyl)-3-methyl-butylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₉H₄₂N₄O₄S, 663.29. found 663.2. -   (7-109)     4′-{6-[(R)-1-(3-aminobenzoylsulfanylmethyl)-3-methyl-butylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₉H₄₂N₄O₄S, 663.29. found 663.2. -   (7-110)     4′-{6-[(R)-1-(4-aminobenzoylsulfanylmethyl)-3-methyl-butylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₉H₄₂N₄O₄S, 663.29. found 663.2. -   (7-111)     4′-{6-[(R)-1-((S)-2-amino-3-methylbutyrylsulfanylmethyl)-3-methylbutyl-carbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₇H₄₆N₄O₄S, 643.32. found 643.4. -   (7-112)     4′-{4-methyl-6-[(R)-3-methyl-1-((S)-pyrrolidine-2-carbonylsulfanylmethyl)-butylcarbamoyl]-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₇H₄₄N₄O₄S, 641.31. found 641.2. -   (7-113)     2-(5-{6-[(R)-1-(2-fluorobenzyl)-2-mercaptoethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}-pyridin-2-yl)benzoic     acid. MS m/z: [M+H⁺] calcd for C₃₄H₃₃FN₄O₃S, 597.23. found 597.6. -   (7-114)     4′-{6-[(R)-1-benzyl-2-(2-dimethylaminoacetylsulfanyl)-ethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₉H₄₂N₄O₄S, 663.29. found 663.2. -   (7-115)     4′-{6-[(R)-1-benzyl-2-(3-dimethylaminopropionylsulfanyl)-ethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₀H₄₄N₄O₄S, 677.31. found 676.2. -   (7-116)     4′-{6-[(R)-1-benzyl-2-(4-dimethylaminobutyrylsulfanyl)-ethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₁H₄₆N₄O₄S, 691.32. found 691.4. -   (7-117)     4′-(6-{(R)-1-benzyl-2-[2-(2-methylpiperidin-1-yl)-acetylsulfanyl]-ethylcarbamoyl}-4-methyl-2-propylbenzoimidazol-1-ylmethyl)biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₃H₄₈N₄O₄S, 717.34. found 717.4. -   (7-118)     4′-(6-{(R)-1-benzyl-2-[2-(3-methylpiperidin-1-yl)-acetylsulfanyl]-ethylcarbamoyl}-4-methyl-2-propylbenzoimidazol-1-ylmethyl)biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₃H₄₈N₄O₄S, 717.34. found 717.4. -   (7-119)     4′-{6-[(R)-1-(4-dimethylaminobutyrylsulfanylmethyl)-3-methylbutyl-carbamoyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₈H₄₈N₄O₄S, 657.34. found 657.4. -   (7-120)     4′-{4-methyl-6-[(R)-3-methyl-1-(2-piperidin-1-yl-acetylsulfanylmethyl)-butylcarbamoyl]-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₉H₄₈N₄O₄S, 669.34. found 669.4. -   (7-121)     4′-{6-[(R)-1-benzyl-2-(3-carboxypropionylsulfanyl)ethylcarbamoyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₉H₃₉N₃O₆S, 678.26. found 678.2. -   (7-122)     4′-{6-[(R)-1-benzyl-2-(4-carboxybenzoylsulfanyl)ethylcarbamoyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₃H₃₉N₃O₆S, 726.26. found 726.2. -   (7-123)     4′-{6-[(R)-2-((S)-2-amino-4-carboxybutyrylsulfanyl)-1-benzylethylcarbamoyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₀H₄₂N₄O₆S, 707.28. found 707.2. -   (7-124)     4′-{6-[(R)-2-((S)-3-amino-3-carboxypropionylsulfanyl)-1-benzylethyl-carbamoyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₉H₄₀N₄O₆S, 693.27. found 693.2. -   (7-125)     4′-{6-[(R)-1-benzyl-2-(3-carboxybenzoylsulfanyl)-ethylcarbamoyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₃H₃₉N₃O₆S, 726.26. found 726.2. -   (7-126)     4′-{6-[(R)-1-(3-carboxypropionylsulfanylmethyl)-3-methylbutylcarbamoyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₆H₄₁N₃O₆S, 644.27. found 644.2. -   (7-127)     4′-{6-[(R)-1-(3-carboxybenzoylsulfanylmethyl)-3-methyl-butylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₀H₄₁N₃O₆S, 692.27. found 692.2. -   (7-128)     4′-{6-[(R)-1-(4-carboxybenzoylsulfanylmethyl)-3-methyl-butylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₀H₄₁N₃O₆S, 692.27. found 692.2. -   (7-129)     4′-{6-[(R)-1-((S)-2-amino-4-carboxybutyrylsulfanylmethyl)-3-methylbutyl-carbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₇H₄₄N₄O₆S, 673.30. found 673.2. -   (7-130)     4′-{6-[(R)-1-((S)-3-amino-3-carboxypropionylsulfanylmethyl)-3-methylbutyl-carbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₆H₄₂N₄O₆S, 659.28. found 659.2. -   (7-131)     4′-{6-[(R)-1-(3-fluorobenzyl)-2-(formylhydroxyamino)ethylcarbamoyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₆H₃₅FN₄O₅, 623.26. found 623.2. -   (7-132)     4′-{6-[(R)-1-(2-fluorobenzyl)-2-(formylhydroxyamino)ethylcarbamoyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₆H₃₅FN₄O₅, 623.26. found 623.2. -   (7-133)     4′-{6-[(R)-2-(formylhydroxyamino)-1-(4-methylbenzyl)ethylcarbamoyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₇H₃₈N₄O₅, 619.28. found 619.2. -   (7-134) 4′-{6-[(R)-2-((S)-2-amino-3-carbo     xypropionylsulfanyl)-1-benzylethyl-carbamoyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₉H₄₀N₄O₆S, 693.27. found 693.2. -   (7-135)     4′-{6-[(R)-2-((S)-4-amino-4-carboxybutyrylsulfanyl)-1-benzylethylcarbamoyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₄₀H₄₂N₄O₆S, 707.28. found 707.6. -   (7-136) 4′-{6-[(R)-1-((S)-4-amino-4-carbo     xybutyrylsulfanylmethyl)-3-methylbutyl-carbamoyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₇H₄₄N₄O₆S, 673.30. found 673.6. -   (7-137)     4′-{6-[(R)-1-((R)-2-amino-3-carboxypropionylsulfanylmethyl)-3-methylbutyl-carbamoyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₆H₄₂N₄O₆S, 659.28. found 659.6. -   (7-138) 4′-{6-[(R)-1-(2-dimethylamino     acetylsulfanylmethyl)-3-methylbutylcarb     amoyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₆H₄₄N₄O₄S, 629.31. found 629.8.

Example 8

Following the procedures described in the examples above, and substituting the appropriate starting materials and reagents, compounds 8-1 to 8-50, having the following formula, were also prepared:

Ex. R R⁵ R⁶ 8-1 H —CH₂—SH —CH₂CH(CH₃)₂ 8-2 H —CH₂—SH benzyl 8-3 H —CH₂—SH —CH₂-cyclopropyl 8-4 H —SH benzyl 8-5 H —C(O)N(OH)H benzyl 8-6 H —CH₂—SH —(CH₂)₃CH₃ 8-7 H —SH —(CH₂)₂CH₃ 8-8 H —SH —(CH₂)₂CH₃ 8-9 H —CH₂—SH —CH(CH₃)₂ 8-10 Br —CH₂—SH —CH₂-cyclohexyl 8-11 Br —SH —CH₂CH(CH₃)₂ 8-12 Br —SH —CH₂C(CH₃)₃ 8-13 Br —SH —(CH₂)₂CF₃ 8-14 Br —CH₂—SH —CH₂CH(CH₃)₂ 8-15 Br —CH₂—SH —CH(CH₃)₂ 8-16 Br —SH —(CH₂)₂CH₃ 8-17 Br —SH benzyl 8-18 Br —CH₂—SH benzyl 8-19 Br —CH₂—SH —(CH₂)₂CH₃ 8-20 Br —CH₂—SH —(CH₂)₂CH(CH₃)₂ 8-21 Br —SH —CH₂CH(CH₃)₂ 8-22 Br —SH —CH₂C(CH₃)₃ 8-23 Br —SH —(CH₂)₂CH₃ 8-24 Br —CH₂—SH —CH₂CH(CH₃)₂ 8-25 H —SH —CH₂CH(CH₃)₂ 8-26 H —SH —CH₂C(CH₃)₃ 8-27 H —SH —(CH₂)₂CF₃ 8-28 H —SH —(CH₂)₃CF₃ 8-29 H —SH —CH₂CH(CH₃)₂ 8-30 H —CH₂—SH —CH₂CH(CH₃)₂ 8-31 H —CH₂—SH —CH₂CH(CH₃)₂ 8-32 H —SH —CH₂C(CH₃)₃ 8-33 Br —CH₂—SH —CH₂CH(CH₃)₂ 8-34 F —CH₂—SH —CH₂CH(CH₃)₂ 8-35 F —SH —CH₂C(CH₃)₃ 8-36 F —SH —(CH₂)₂CF₃ 8-37 F —SH —(CH₂)₃CF₃ 8-38 F —SH —(CH₂)₂CH₃ 8-39 F —CH₂—SH —(CH₂)₃CH₃ 8-40 F —CH₂—SH —(CH₂)₂CH(CH₃)₂ 8-41 F —CH₂—SH —CH(CH₃)CH₂CH₃ 8-42 F —CH₂—SH —CH₂-cyclopentyl 8-43 F —CH₂—SH —CH₂-cyclohexyl 8-44 F —CH₂—SH benzyl 8-45 F —SH benzyl 8-46 F —CH₂—SH —CH₂CH(CH₃)₂ 8-47 F —CH₂—SH —CH(CH₃)₂ 8-48 F —SH —(CH₂)₂CH₃ 8-49 F —SH —CH₂C(CH₃)₃ 8-50 F —SH —CH₂CH(CH₃)₂

-   (8-1)     4-{6-[(2-mercaptomethyl-4-methylpentanoylamino)methyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₇H₃₅N₃O₃S, 482.24. found 482.2. -   (8-2)     4-{6-[(2-mercaptomethyl-3-phenylpropionylamino)methyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₃₀H₃₃N₃O₃S, 516.22. found 516.2. -   (8-3)     4-{6-[(3-cyclopropyl-2-mercaptomethyl-propionylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₇H₃₃N₃O₃S, 480.22. found 480.2. -   (8-4)     4-{6-[((S)-2-mercapto-3-phenylpropionylamino)methyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₉H₃₁N₃O₃S, 502.21. found 502.2. -   (8-5)     4-{6-[(2-hydroxycarbamoyl-3-phenylpropionylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₃₀H₃₂N₄O₅, 529.24. found 529.2. -   (8-6)     4-{6-[(2-mercaptomethyl-hexanoylamino)methyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₇H₃₅N₃O₃S, 482.24. found 482.2. -   (8-7)     4-{6-[((R)-2-mercaptopentanoylamino)methyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₅H₃₁N₃O₃S, 454.21. found 454.2. -   (8-8)     4-{6-[((S)-2-mercaptopentanoylamino)methyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₅H₃₁N₃O₃S, 454.21. found 454.2. -   (8-9)     4-{6-[((S)-2-mercaptomethyl-3-methyl-butyrylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₆H₃₃N₃O₃S, 468.22. found 468.2. -   (8-10)     2-bromo-4-{6-[(3-cyclohexyl-2-mercaptomethyl-propionylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₃₀H₃₈BrN₃O₃S, 600.18. found 601.2. -   (8-11)     2-bromo-4-{6-[((S)-2-mercapto-4-methylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₆H₃₂BrN₃O₃S, 546.13. found 547.2. -   (8-12)     2-bromo-4-{6-[((S)-2-mercapto-4,4-dimethylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₇H₃₄BrN₃O₃S, 560.15. found 561.2. -   (8-13)     2-bromo-4-{4-methyl-2-propyl-6-[(5,5,5-trifluoro-2-mercaptopentanoyl-amino)methyl]-benzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₅H₂₇BrF₃N₃O₃S, 586.09. found 587.2. -   (8-14)     2-bromo-4-{6-[((S)-2-mercaptomethyl-4-methylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₇H₃₄BrN₃O₃S, 560.15. found 561.2. -   (8-15)     2-bromo-4-{6-[((S)-2-mercaptomethyl-3-methyl-butyrylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₆H₃₂BrN₃O₃S, 546.13. found 547.2. -   (8-16)     2-bromo-4-{6-[((S)-2-mercaptopentanoylamino)methyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₅H₃₀BrN₃O₃S, 532.12. found 533.2. -   (8-17)     2-bromo-4-{6-[((S)-2-mercapto-3-phenylpropionylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₉H₃₀BrN₃O₃S, 580.12. found 581.2. -   (8-18)     2-bromo-4-{6-[(2-mercaptomethyl-3-phenylpropionylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₃₀H₃₂BrN₃O₃S, 594.13. found 595.2. -   (8-19)     2-bromo-4-{6-[(2-mercaptomethylhexanoylamino)methyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₇H₃₄BrN₃O₃S, 560.15. found 561.2. -   (8-20)     2-bromo-4-{6-[(2-mercaptomethyl-5-methylhexanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₈H₃₆BrN₃O₃S, 574.17. found 575.2. -   (8-21)     2-bromo-4-{6-[((R)-2-mercapto-4-methylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₆H₃₂BrN₃O₃S, 546.13. found 547.2. -   (8-22)     2-bromo-4-{6-[((R)-2-mercapto-4,4-dimethylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₇H₃₄BrN₃O₃S, 560.15. found 561.2. -   (8-23)     2-bromo-4-{6-[((R)-2-mercaptopentanoylamino)methyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₅H₃₀BrN₃O₃S , 532.12. found 533.2. -   (8-24)     2-bromo-4-{6-[((R)-2-mercaptomethyl-4-methylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₇H₃₄BrN₃O₃S, 560.15. found 561.2. -   (8-25)     4-{6-[((S)-2-mercapto-4-methylpentanoylamino)methyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₆H₃₃N₃O₃S, 468.22. found 468.2. -   (8-26)     4-{6-[((S)-2-mercapto-4,4-dimethylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₇H₃₅N₃O₃S, 482.24. found 482.2. -   (8-27)     4-{4-methyl-2-propyl-6-[(5,5,5-trifluoro-2-mercaptopentanoylamino)methyl]-benzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₅H₂₈F₃N₃O₃S, 508.18. found 508.2. -   (8-28)     4-{4-methyl-2-propyl-6-[(6,6,6-trifluoro-2-mercaptohexanoylamino)methyl]-benzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₆H₃₀F₃N₃O₃S, 522.20. found 522.2. -   (8-29)     4-{6-[((R)-2-mercapto-4-methylpentanoylamino)methyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₆H₃₃N₃O₃S, 468.22. found 468.2. -   (8-30)     4-{6-[((R)-2-mercaptomethyl-4-methylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₇H₃₅N₃O₃S, 482.24. found 482.2. -   (8-31)     4-{6-[((S)-2-mercaptomethyl-4-methylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₇H₃₅N₃O₃S, 482.24. found 482.2. -   (8-32)     4-{6-[((R)-2-mercapto-4,4-dimethylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₇H₃₅N₃O₃S, 482.24. found 482.2. -   (8-33)     2-bromo-4-{6-[(2-mercaptomethyl-4-methylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₇H₃₄BrN₃O₃S, 560.15. found 561.2. -   (8-34)     2-fluoro-4-{6-[((S)-2-mercaptomethyl-4-methylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₇H₃₄FN₃O₃S, 500.23. found 500.2. -   (8-35)     2-fluoro-4-{6-[((S)-2-mercapto-4,4-dimethylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₇H₃₄FN₃O₃S, 500.23. found 500.2. -   (8-36)     2-fluoro-4-{4-methyl-2-propyl-6-[(5,5,5-trifluoro-2-mercaptopentanoylamino)-methyl]benzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₅H₂₇F₄N₃O₃S, 526.17. found 526.2. -   (8-37)     2-fluoro-4-{4-methyl-2-propyl-6-[(6,6,6-trifluoro-2-mercaptohexanoylamino)-methyl]benzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₆H₂₉F₄N₃O₃S, 540.19. found 540.2. -   (8-38)     2-fluoro-4-{6-[((S)-2-mercaptopentanoylamino)methyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₅H₃₀FN₃O₃S, 472.20. found 472.2. -   (8-39)     2-fluoro-4-{6-[(2-mercaptomethyl-hexanoylamino)methyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₇H₃₄FN₃O₃S, 500.23. found 500.2. -   (8-40)     2-fluoro-4-{6-[(2-mercaptomethyl-5-methyl-hexanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₈H₃₆FN₃O₃S, 514.25. found 514.2. -   (8-41)     2-fluoro-4-{6-[(2-mercaptomethyl-3-methylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₇H₃₄FN₃O₃S, 500.23. found 500.2. -   (8-42)     4-{6-[(3-cyclopentyl-2-mercaptomethyl-propionylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}-2-fluorobenzoic     acid. MS m/z: [M+H⁺] calcd for C₂₉H₃₆FN₃O₃S, 526.25. found 526.2. -   (8-43)     4-{6-[(3-cyclohexyl-2-mercaptomethyl-propionylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}-2-fluorobenzoic     acid. MS m/z: [M+H⁺] calcd for C₃₀H₃₈FN₃O₃S, 540.26. found 540.2. -   (8-44)     2-fluoro-4-{6-[(2-mercaptomethyl-3-phenylpropionylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₃₀H₃₂FN₃O₃S, 534.22. found 534.2. -   (8-45)     2-fluoro-4-{6-[((S)-2-mercapto-3-phenylpropionylamino)methyl}-4-methyl-2-propylbenzoimidazol-1-ylmethyl]benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₉H₃₀FN₃O₃S, 520.20. found 520.2. -   (8-46)     2-fluoro-4-{6-[((R)-2-mercaptomethyl-4-methylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₇H₃₄FN₃O₃S, 500.23. found 500.2. -   (8-47)     2-fluoro-4-{6-[((S)-2-mercaptomethyl-3-methyl-butyrylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₆H₃₂FN₃O₃S, 486.22. found 486.2. -   (8-48)     2-fluoro-4-{6-[((R)-2-mercaptopentanoylamino)methyl]-4-methyl-2-propyl-benzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₅H₃₀FN₃O₃S, 472.20. found 472.2. -   (8-49)     2-fluoro-4-{6-[((R)-2-mercapto-4,4-dimethylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₇H₃₄FN₃O₃S, 500.23. found 500.2. -   (8-50)     2-fluoro-4-{6-[((R)-2-mercapto-4-methylpentanoylamino)methyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₆H₃₂FN₃O₃S, 486.22. found 486.2.

Example 9

Following the procedures described in the examples above, and substituting the appropriate starting materials and reagents, compounds 9-1 to 9-6, having the following formula, were also prepared:

Ex. R¹ R⁵ R⁶ 9-1 —COOH —CH₂—C(O)N(OH)H benzyl 9-2 —COOH —CH₂—N(OH)C(O)H benzyl 9-3 —COOH —CH₂—N(OH)C(O)CH₃ benzyl 9-4 —COOCH₃ —CH₂—N(OH)C(O)H benzyl 9-5 —COOH —CH₂SH —CH₂CH(CH₃)₂ 9-6 —COOH —CH₂SH benzyl

-   (9-1)     4-[6-((R)-1-benzyl-2-hydroxycarbamoylethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]benzoic     acid. MS m/z: [M+H⁺] calcd for C₃₀H₃₂N₄O₅, 529.24. found 529.6. -   (9-2)     4-{6-[(R)-1-benzyl-2-(formylhydroxyamino)ethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₃₀H₃₂N₄O₅, 529.24. found 529.5. -   (9-3) 4-{6-[(R)-2-(ac     etylhydroxyamino)-1-benzylethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₃₁H₃₄N₄O₅, 543.25. found 543.2. -   (9-4)     4-{6-[(R)-1-benzyl-2-(formylhydroxyamino)ethylcarbamoyl]-4-methyl-2-propylbenzoimidazol-1-ylmethyl}benzoic     acid methyl ester. MS m/z: [M+H⁺] calcd for C₃₁H₃₄N₄O₅, 543.25.     found 543.3. -   (9-5)     4-[6-((R)-1-mercaptomethyl-3-methylbutylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]-benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₆H₃₃N₃O₃S, 468.22. found 468.2. -   (9-6) 4-[6-((R)-1-benzyl-2-mercapto     ethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]-benzoic     acid. MS m/z: [M+H⁺] calcd for C₂₉H₃₁N₃O₃S, 502.21. found 502.4.

Preparation 11 Dimer of 7-Methyl-2-propyl-3H-benzoimidazole-5-carboxylic acid ((R)-1-mercaptomethyl-3-methylbutyl)amide

7-Methyl-2-propyl-3H-benzoimidazole-5-carboxylic acid (10 g, 40 mmol) and HATU (19.2 g, 50.4 mmol) were dissolved in DMF (21.4 mL, 276.7 mmol), and stirred for 5 minutes at room temperature. (R)-1-((R)-2-Amino-4-methylpentyldisulfanyl-methyl)-3-methylbutylamine.2[HC1] (7.7 g, 22.9 mmol) was added, followed by DIPEA (18.0 mL, 103 mmol). The mixture was stirred at room temperature overnight. EtOAc (200 mL) was added and the mixture was washed with a saturated sodium bicarbonate solution (100 mL) and saturated aqueous NaCl (2×100 mL). The organic layer was dried over MgSO₄, filtered, and evacuated to dryness. The reaction mixture was concentrated and the resulting crude solid was purified by silica gel chromatography.

Preparation 12 4′-Bromomethylbiphenyl-2-sulfonic Acid 1-dimethylaminometh-(E)-ylideneamide

2-Bromo-N[1-dimethylaminometh-(E)-ylidene]benzenesulfonamide

1,1-Dimethoxy-N,N-dimethylmethanamine (14.6 mL, 104 mmol) was added to 2-bromobenzene-1-sulfonamide (20.4 g, 86.4 mmol) in DMF (56 mL, 720 mmol). The mixture was stirred at room temperature for about 90 minutes. A solution of sodium hydrogen sulfate (1.7 g, 14 mmol) in water (170 mL, 9400 mmol) was cooled to 0° C. and then added to the mixture. The precipitate was filtered, washed with water, and dried to yield 2-bromo-N[1-dimethylaminometh-(E)-ylidene]benzenesulfonamide as a white solid (24.3 g).

4′-Methybiphenyl-2-sulfonic acid 1-dimethylaminometh-(E)-ylideneamide

2-Bromo-N-[1-dimethylaminometh-(E)-ylidene]benzenesulfonamide (5.36 g, 18.4 mmol), 4-methylphenylboronic acid (5.0 g, 36.8 mmol), potassium carbonate (5.1 g, 36.8 mmol), water (19.7 mL, 1090 mmol), EtOH (49.2 mL, 842 mmol) and toluene (98.3 mL, 923 mmol) were combined and the mixture was stirred under nitrogen. Tetrakis(triphenylphosphine)palladium(0) (1.4 g, 1.2 mmol) was then added. The mixture was heated at 60° C. for about 2 hours, then heated at 70° C. for 30 minutes. The mixture was then cooled to room temperature and water (100 mL) and EtOAc (100 mL) were added. The mixture was washed with saturated aqueous NaCl, extracted with EtOAc, dried over MgSO₄, filtered through Celite® and concentrated to yield the crude product as a red solid (8.6 g). The solid material was triturated with 1:1 EtOAc:hexanes, filtered, and rinsed with hexanes to yield the crude product as a reddish-brown solid (6.2 g). The solid material was again triturated with EtOAc, filtered, and rinsed with EtOAc to yield 4′-methybiphenyl-2-sulfonic acid 1-dimethylaminometh-(E)-ylideneamide as a light brown solid (4.6 g; 96.2% purity).

Title Compound

4′-Methylbiphenyl-2-sulfonic acid 1-dimethylaminometh-(E)-ylideneamide (7.8 g, 26 mmol) was combined with NBS (4.6 g, 25.8 mmol), benzoyl peroxide (31 mg, 130 mmol) and chlorobenzene (100 mL, 990 mmol). The mixture was heated at 95° C. The reaction was monitored until completion (about 2.5 hours). The mixture was then cooled to room temperature and water was added. The mixture was extracted with DCM, washed with a saturated bicarbonate solution and saturated aqueous NaCl, extracted again with DCM, dried over MgSO₄, filtered, and concentrated. The crude material (9.8 g) was combined with a second lot of crude material (1.7 g) and taken up in DCM (40 mL). EtOAc (120 mL) was added. A few seed crystals from a third lot of material were added and the mixture was stored at −20° C. A precipitate formed, was filtered and rinsed with EtOAc. HPLC analysis of the resulting product, an off-white solid (8.7 g) showed 17% starting material, 74% title compound, 7% dibromide.

Example 10 3-(2′-Acetylsulfamoylbiphenyl-4-ylmethyl)-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic acid ((R)-1-mercaptomethyl-3-methylbutyl)amide

Dimer of 3-(2′-{[1-dimethylamino-meth-(E)-ylidene]-sulfamoyl}biphenyl-4-ylmethyl)-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic acid ((R)-1-mercaptomethyl-3-methylbutyl)amide (10a): The 7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic acid ((R)-1-mercaptomethyl-3-methylbutyl)amide dimer (2.5 g, 3.7 mmol) was dissolved in DMF (50.0 mL, 646 mmol). The mixture was cooled to 0° C., and sodium hydride (356 mg, 8.89 mmol) was added in portions. 4′-Bromomethylbiphenyl-2-sulfonic acid 1-dimethylaminometh-(E)-ylideneamide (4.0 g, 7.8 mmol) was added and the mixture was stirred at 0° C. for 10 minutes, then warmed to room temperature. Water was added upon completion of the reaction (about 40 minutes). The material was extracted 3× with DCM, washed with saturated aqueous NaCl, dried over MgSO₄, filtered, and concentrated to yield 6.1 g of crude intermediate 10a.

Dimer of 7-methyl-2-propyl-3-(2′-sulfamoylbiphenyl-4-ylmethyl)-3H-benzoimidazole-5-carboxylic acid ((R)-1-mercaptomethyl-3-methylbutyl)amide (10b): Intermediate 10a was taken up in EtOH (110 mL, 1800 mmol). Concentrated HCl (55.0 mL) was added and the mixture was heated at 70° C. for 4 hours, and at 75° C. for 30 minutes. The mixture was then cooled to room temperature. 6N NaOH (110 mL) was added to pH 13. Concentrated HCl (2 mL) was then added, maintaining the pH 13. The mixture was extracted with 3:1 CHCl₃:iPrOH (3×), washed with saturated aqueous NaCl, extracted again with 3:1 CHCl₃:iPrOH, dried over MgSO₄, filtered, and concentrated to yield a light brown solid (4.8 g). The solid was taken up in EtOAc and a small amount of MeOH, filtered and rinsed with EtOAc to yield a chunky off-white solid (1.8 g). The liquor and the solid were combined and taken up in 3:1 CHCl₃:iPrOH and water. 1N HCl was added until pH 1. Saturated aqueous bicarbonate was added until pH 9. The product was extracted with 3:1 CHCl₃:iPrOH (3×), washed with saturated aqueous NaCl, dried over MgSO₄, filtered, and concentrated to yield a yellow solid (4.3 g); this solid was taken up in EtOAc and a small amount of MeOH, filtered and rinsed with EtOAc to yield intermediate 10b as a powdery off-white solid (1.4 g). The liquor was concentrated to yield a yellow solid (2.6 g), which was purified by silica gel chromatography (120g, 0% to 10% to 40% MeOH in DCM) to yield a yellow solid (1.3 g); this solid was taken up in EtOAc and a small amount of MeOH, filtered and rinsed with EtOAc to yield intermediate 10b as an off-white solid (0.5 g).

Dimer of 3-(2′-acetylsulfamoylbiphenyl-4-ylmethyl)-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic acid ((R)-1-mercaptomethyl-3-methylbutyl)amide (10c): Intermediate 10b (2 g, 1.7 mmol) was taken up in DCM (50.0 mL, 780 mmol) and THF (25.0 mL, 308 mmol). DIPEA (1480 μL, 8.48 mmol) was added, followed by the addition of 4-dimethylaminopyridine (518 mg, 4.2 mmol) and acetyl chloride (724 μL, 10.2 mmol). The mixture was stirred at room temperature for 10 minutes. Additional DIPEA (592 μL, 2 eq) was added, followed by additional acetyl chloride (121 μL, 1 eq) and 4-dimethylaminopyridine (52 mg, 0.25 eq). Water (50 mL) was added after 10 minutes. The mixture was extracted with DCM (2×), washed with saturated aqueous NaCl, dried over MgSO₄, filtered, and concentrated to yield crude intermediate 10c (3.0 g).

Intermediate 10c (3.0 g) was dissolved in THF (80 mL), and tris(2-carboxyethyl)phosphine hydrochloride (1460 mg, 5.1 mmol) in water (40 mL) was added. The mixture was stirred under nitrogen for 40 minutes. The mixture was extracted with DCM (3×), washed with saturated aqueous NaCl, dried over MgSO₄, filtered, concentrated, and purified by preparative HPLC to yield the title compound as a white solid (1.3 g; 99.6% purity). MS m/z: [M+H⁺] calcd for C₃₃H₄₀N₄O₄S₂, 621.25. found 621.6.

Example 11

Following the procedures described in the examples above, and substituting the appropriate starting materials and reagents, compounds II-1 to 11-16, having the following formula, were also prepared:

Ex. R¹ R⁵ R⁶ 11-1 —SO₂NHC(O)-phenyl —CH₂SH benzyl 11-2 —SO₂NHC(O)CH₃ —CH₂SH benzyl 11-3 —SO₂NHC(O)NHCH₂CH₃ —CH₂SH benzyl 11-4 —SO₂NHC(O)(CH₂)₂CH₃ —CH₂SH benzyl 11-5 —SO₂NHC(O)CH₂OH —CH₂SH benzyl 11-6 —SO₂NHC(O)CH₃ —CH₂N(OH)—CHO benzyl 11-7 —SO₂NHC(O)CH₂OH —CH₂SH —CH₂CH(CH₃)₂ 11-8 —SO₂NHC(O)CH(CH₃)(OH) —CH₂SH —CH₂CH(CH₃)₂ 11-9 —SO₂NHC(O)C(CH₃)₂(OH) —CH₂SH —CH₂CH(CH₃)₂ 11-10 —SO₂NHC(O)CH₂OCH₃ —CH₂SH —CH₂CH(CH₃)₂ 11-11 —SO₂NHC(O)CH₂O(CH₂)₂OCH₃ —CH₂SH —CH₂CH(CH₃)₂ 11-12 —SO₂NHC(O)CH₂N(CH₃)₂ —CH₂SH —CH₂CH(CH₃)₂ 11-13 —SO₂NHC(O)CH₂NH₂ —CH₂SH —CH₂CH(CH₃)₂ 11-14 —SO₂NHC(O)CH(CH₃)(NH₂) —CH₂SH —CH₂CH(CH₃)₂ 11-15 —SO₂NHC(O)CH₂-morphohn-4-yl —CH₂SH —CH₂CH(CH₃)₂ 11-16 —SO₂NHC(O)CH₂-4-acetyl- —CH₂SH —CH₂CH(CH₃)₂ piperazin-1-yl

-   (11-1)     3-(2′-benzoylsulfamoylbiphenyl-4-ylmethyl)-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic     acid ((R)-1-benzyl-2-mercaptoethyl)amide. MS m/z: [M+H⁺] calcd for     C₄₁H₄₀N₄O₄S₂, 717.25. found 717.5. -   (11-2)     3-(2′-acetylsulfamoylbiphenyl-4-ylmethyl)-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic     acid ((R)-1-benzyl-2-mercaptoethyl)amide. MS m/z: [M+H⁺] calcd for     C₃₆H₃₈N₄O₄S₂, 655.23. found 655.6. -   (11-3) MS m/z: [M+H⁺] calcd for C₃₇H₄₁N₅O₄S₂, 684.26. found 684.6. -   (11-4)     3-(2′-butyrylsulfamoylbiphenyl-4-ylmethyl)-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic     acid ((R)-1-benzyl-2-mercapto-ethyl)amide. MS m/z: [M+H⁺] calcd for     C₃₈H₄₂N₄O₄S₂, 683.27. found 683.6. -   (11-5)     3-[2′-(2-hydroxyacetylsulfamoyl)-biphenyl-4-ylmethyl]-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic     acid ((R)-1-benzyl-2-mercapto-ethyl)amide. MS m/z: [M+H⁺] calcd for     C₃₆H₃₈N₄O₅S₂, 671.23. found 671.6. -   (11-6)     3-(2′-acetylsulfamoylbiphenyl-4-ylmethyl)-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic     acid [(R)-1-benzyl-2-(formylhydroxyamino)ethyl]amide. MS m/z: [M+H⁺]     calcd for C₃₇H₃₉N₅O₆S, 682.26. found 682.6. -   (11-7)     3-[2′-(2-hydroxyacetylsulfamoyl)-biphenyl-4-ylmethyl]-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic     acid ((R)-1-mercaptomethyl-3-methylbutyl)amide. MS m/z: [M+H⁺] calcd     for C₃₃H₄₀N₄O₅S₂, 637.24. found 637.4. -   (11-8)     3-[2′-((S)-2-hydroxypropionylsulfamoyl)-biphenyl-4-ylmethyl]-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic     acid ((R)-1-mercaptomethyl-3-methylbutyl)amide. MS m/z: [M+H⁺] calcd     for C₃₄H₄₂N₄O₅S₂, 651.26. found 651.2. -   (11-9)     3-[2′-(2-hydroxy-2-methylpropionylsulfamoyl)-biphenyl-4-ylmethyl]-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic     acid ((R)-1-mercaptomethyl-3-methylbutyl)-amide. MS m/z: [M+H⁺]     calcd for C₃₅H₄₄N₄O₅S₂, 665.28. found 665.2. -   (11-10)     3-[2′-(2-methoxyacetylsulfamoyl)biphenyl-4-ylmethyl]-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic     acid ((R)-1-mercaptomethyl-3-methylbutyl)amide. MS m/z: [M+H⁺] calcd     for C₃₄H₄₂N₄O₅S₂, 651.26. found 651.4. -   (11-11)     3-{2′-[2-(2-methoxyethoxy-acetylsulfamoyl]-biphenyl-4-ylmethyl}-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic     acid ((R)-1-mercaptomethyl-3-methylbutyl)-amide. MS m/z: [M+H⁺]     calcd for C₃₆H₄₆N₄O₆S₂, 695.29. found 695.6. -   (11-12) 3-[2′-(2-dimethylamino ac     etylsulfamoyl)-biphenyl-4-ylmethyl]-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic     acid ((R)-1-mercaptomethyl-3-methylbutyl)-amide. MS m/z: [M+H⁺]     calcd for C₃₅H₄₅N₅O₄S₂, 664.29. found 664.4. -   (11-13)     3-[2′-(2-aminoacetylsulfamoyl)-biphenyl-4-ylmethyl]-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic     acid ((R)-1-mercaptomethyl-3-methylbutyl)-amide. MS m/z: [M+H⁺]     calcd for C₃₃H₄₁N₅O₄S₂, 636.26. found 636.2. -   (11-14)     3-[2′4(S)-2-aminopropionylsulfamoyl)-biphenyl-4-ylmethyl]-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic     acid ((R)-1-mercaptomethyl-3-methylbutyl)-amide. MS m/z: [M+H⁺]     calcd for C₃₄H₄₃N₅O₄S₂, 650.28. found 650.6. -   (11-15)     7-methyl-3-[2′-(2-morpholin-4-yl-acetylsulfamoyl)-biphenyl-4-ylmethyl]-2-propyl-3H-benzoimidazole-5-carboxylic     acid ((R)-1-mercaptomethyl-3-methylbutyl)-amide. MS m/z: [M+H⁺]     calcd for C₃₇H₄₇N₅O₅S₂, 706.30. found 706.8. -   (11-16)     3-{2′-[2-(4-acetylpiperazin-1-yl)-acetylsulfamoyl]-biphenyl-4-ylmethyl}-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic     acid ((R)-1-mercaptomethyl-3-methyl-butyl)-amide. MS m/z: [M+H⁺]     calcd for C₃₉H₅₀N₆O₅S₂, 747.33. found 747.6.

Example 12

Following the procedures described in the examples above, and substituting the appropriate starting materials and reagents, compounds 12-1 to 12-17, having the following formula, were also prepared:

Ex. R¹ R² R³ R⁵ R⁶ 12-1 —COOH absent —(CH₂)₂CH₃ —CH₂SH benzyl 12-2 —COOH absent —(CH₂)₂CH₃ —CH₂SH —CH₂CH(CH₃)₂ 12-3 —COOH absent —OCH₂CH₃ —CH₂SH benzyl 12-4 —COOH Cl —(CH₂)₂CH₃ —CH₂SH benzyl 12-5 —COOH Cl —(CH₂)₂CH₃ —CH₂SH —CH₂CH(CH₃)₂ 12-6 —COOH —CF₃ —(CH₂)₂CH₃ —CH₂SH benzyl 12-7 tetrazol-5-yl —CF₃ —(CH₂)₂CH₃ —CH₂SH benzyl 12-8 tetrazol-5-yl —CF₃ —(CH₂)₂CH₃ —CH₂SH —CH₂CH(CH₃)₂ 12-9 tetrazol-5-yl —CF₃ —(CH₂)₂CH₃ —CH₂—N(OH)CHO benzyl 12-10 tetrazol-5-yl Cl —(CH₂)₂CH₃ —CH₂SH benzyl 12-11 tetrazol-5-yl Cl —(CH₂)₂CH₃ —CH₂SH —CH₂CH(CH₃)₂ 12-12 tetrazol-5-yl Cl —(CH₂)₂CH₃ —CH₂—(OH)CHO benzyl 12-13 tetrazol-5-yl Cl —(CH₂)₂CH₃ —CH₂—N(OH)CHO —CH₂CH(CH₃)₂ 12-14 —COOH —CH₂F —(CH₂)₂CH₃ —CH₂SH benzyl 12-15 —COOH —CH₂F —(CH₂)₂CH₃ —CH₂SH —CH₂CH(CH₃)₂ 12-16 —COOH —CH₂OH —(CH₂)₂CH₃ —CH₂SH benzyl 12-17 —COOH —CH₂OH —(CH₂)₂CH₃ —CH₂SH —CH₂CH(CH₃)₂

-   (12-1)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₄H₃₃N₃O₃S, 564.22. found 564.6. -   (12-2)     4′-[6-((R)-1-mercaptomethyl-3-methyl-butylcarbamoyl)-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₁H₃₅N₃O₃S, 530.24. found 530.6. -   (12-3)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-2-ethoxybenzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₃H₃₁N₃O₄S, 566.20. found 566.5. -   (12-4)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-chloro-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₄H₃₂ClN₃O₃S, 598.19. found 598.6. -   (12-5)     4′-[4-chloro-6-((R)-1-mercaptomethyl-3-methylbutylcarbamoyl)-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₁H₃₄ClN₃O₃S, 564.20. found 564.2. -   (12-6)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-2-propyl-4-trifluoromethyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₅H₃₂F₃N₃O₃S, 632.21. found 632.4. -   (12-7)     2-propyl-3-[2′-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]-7-trifluoromethyl-3H-benzoimidazole-5-carboxylic     acid ((R)-1-benzyl-2-mercaptoethyl)amide. MS m/z: [M+H⁺] calcd for     C₃₅H₃₂F₃N₇OS, 656.23. found 656.6. -   (12-8)     2-propyl-3-[2′-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]-7-trifluoromethyl-3H-benzoimidazole-5-carboxylic     acid ((R)-1-mercaptomethyl-3-methylbutyl)amide. MS m/z: [M+H⁺] calcd     for C₃₂H₃₄F₃N₇OS, 622.25. found 622.6. -   (12-9)     2-propyl-3-[2′-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]-7-trifluoromethyl-3H-benzoimidazole-5-carboxylic     acid [(R)-1-benzyl-2-(formylhydroxyamino)ethyl]amide. MS m/z: [M+H⁺]     calcd for C₃₆H₃₃F₃N₈O₃, 683.26. found 683.6. -   (12-10)     7-chloro-2-propyl-3-[2′-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]-3H-benzoimidazole-5-carboxylic     acid ((R)-1-benzyl-2-mercaptoethyl)amide. MS m/z: [M+H⁺] calcd for     C₃₄H₃₂ClN₇OS, 622.21. found 622.5. -   (12-11)     7-chloro-2-propyl-3-[2′-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]-3H-benzoimidazole-5-carboxylic     acid ((R)-1-mercaptomethyl-3-methylbutyl)amide. MS m/z: [M+H⁺] calcd     for C₃₁H₃₄ClN₇OS, 588.22. found 588.6. -   (12-12)     7-chloro-2-propyl-3-[2′-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]-3H-benzoimidazole-5-carboxylic     acid [(R)-1-benzyl-2-(formylhydroxyamino)ethyl]amide. MS m/z: [M+H⁺]     calcd for C₃₅H₃₃ClN₈O₃, 649.24. found 649.8. -   (12-13)     7-chloro-2-propyl-3-[2′-(1H-tetrazol-5-yl)-biphenyl-4-ylmethyl]-3H-benzoimidazole-5-carboxylic     acid {(R)-1-[(formylhydroxyamino)methyl]-3-methylbutyl}amide. MS     m/z: [M+H⁺] calcd for C₃₂H₃₅ClN₈O₃, 615.25. found 615.4. -   (12-14)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-fluoromethyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₅H₃₄FN₃O₃S, 596.23. found 596.6. -   (12-15)     4′-[4-fluoromethyl-6-((R)-1-mercaptomethyl-3-methyl-butylcarbamoyl)-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₂H₃₆FN₃O₃S, 562.25. found 562.4. -   (12-16)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-hydroxymethyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₅H₃₅N₃O₄S, 594.24. found 594.4. -   (12-17)     4′-[4-hydroxymethyl-6-((R)-1-mercaptomethyl-3-methylbutylcarbamoyl)-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₂H₃₇N₃O₄S, 560.25. found 560.4.

Example 13

Following the procedures described in the examples above, and substituting the appropriate starting materials and reagents, compounds 13-1 to 13-8, having the following formula, were also prepared:

Ex. R R⁶ 13-1 2′-Cl benzyl 13-2 2′-F benzyl 13-3 3′-Cl benzyl 13-4 3′-F benzyl 13-5 2′-F —CH₂CH(CH₃)₂ 13-6 2′-Cl —CH₂CH(CH₃)₂ 13-7 3′-Cl —CH₂CH(CH₃)₂ 13-8 3′-F —CH₂CH(CH₃)₂

-   (13-1)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]-2′-chlorobiphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₅H₃₄ClN₃O₃S, 612.20. found 612.2. -   (13-2)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]-2′-fluorobiphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₅H₃₄FN₃O₃S, 596.23. found 596.2. -   (13-3)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]-3′-chlorobiphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₅H₃₄ClN₃O₃S, 612.20. found 612.2. -   (13-4)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]-3′-fluorobiphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₅H₃₄FN₃O₃S, 596.23. found 596.2. -   (13-5)     2′-fluoro-4′-[6-((R)-1-mercaptomethyl-3-methylbutylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₂H₃₆FN₃O₃S, 562.25. found 562.2. -   (13-6)     2′-chloro-4′-[6-((R)-1-mercaptomethyl-3-methylbutylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₂H₃₆ClN₃O₃S, 578.22. found 578.2. -   (13-7)     3′-chloro-4′-[6-((R)-1-mercaptomethyl-3-methylbutylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₂H₃₆ClN₃O₃S, 578.22. found 578.2. -   (13-8)     3′-fluoro-4′-[6-((R)-1-mercaptomethyl-3-methylbutylcarbamoyl)-4-methyl-2-propylbenzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₂H₃₆FN₃O₃S, 562.25. found 562.2.

Example 14

Following the procedures described in the examples above, and substituting the appropriate starting materials and reagents, compounds 14-1 to 14-7, having the following formula, were also prepared:

Ex. R R⁶ 14-1 3-Cl benzyl 14-2 4-Cl benzyl 14-3 5-Cl benzyl 14-4 6-Cl benzyl 14-5 3-Cl —CH₂CH(CH₃)₂ 14-6 4-Cl —CH₂CH(CH₃)₂ 14-7 5-Cl —CH₂CH(CH₃)₂

-   (14-1)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]-3-chlorobiphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₅H₃₄ClN₃O₃S, ; 612.20 found 612.2. -   (14-2)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]-4-chlorobiphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₅H₃₄ClN₃O₃S, 612.20. found 612.2. -   (14-3)     4′-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]-5-chlorobiphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₅H₃₄ClN₃O₃S, 612.20. found 612.2. -   (14-4)     4′-[6-((R)-1-benzyl-2-mercapt-ethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]-6-chlorobiphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₅H₃₄ClN₃O₃S, 612.20. found 612.2. -   (14-5)     3-chloro-4′-[6-((R)-1-mercaptomethyl-3-methylbutylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]-biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₂H₃₆ClN₃O₃S, 578.22. found 578.2. -   (14-6)     4-chloro-4′-[6-((R)-1-mercaptomethyl-3-methylbutylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₂H₃₆ClN₃O₃S, 578.22. found 578.2. -   (14-7)     5-chloro-4′-[6-((R)-1-mercaptomethyl-3-methylbutylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]biphenyl-2-carboxylic     acid. MS m/z: [M+H⁺] calcd for C₃₂H₃₆ClN₃O₃S, 578.22. found 578.2.

Example 15

Following the procedures described in the examples above, and substituting the appropriate starting materials and reagents, compounds 15-1 to 15-4, having the following formula, were also prepared:

Ex. R¹ R⁶ 15-1 —COOH benzyl 15-2 —COOH —CH₂CH(CH₃)₂ 15-3 —SO₂NHC(O)CH₃ benzyl 15-4 —SO₂NHC(O)CH₃ —CH₂CH(CH₃)₂

-   (15-1)     2-{5-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]-pyridin-2-yl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₃₄H₃₄N₄O₃S, 579.24. found 579.5. -   (15-2)     2-{5-[6-((R)-1-mercaptomethyl-3-methylbutylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]-pyridin-2-yl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₃₁H₃₆N₄O₃S, 545.25. found 545.5. -   (15-3)     3-[6-(2-acetylsulfamoylphenyl)-pyridin-3-ylmethyl]-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic     acid ((R)-1-benzyl-2-mercaptoethyl)amide. MS m/z: [M+H⁺] calcd for     C₃₅H₃₇N₅O₄S₂, 656.23. found 656.4. -   (15-4)     3-[6-(2-acetylsulfamoylphenyl)-pyridin-3-ylmethyl]-7-methyl-2-propyl-3H-benzoimidazole-5-carboxylic     acid ((R)-1-mercaptomethyl-3-methylbutyl)amide. MS m/z: [M+H⁺] calcd     for C₃₂H₃₉N₅O₄S₂, 622.24. found 622.5.

Example 16

Following the procedures described in the examples above, and substituting the appropriate starting materials and reagents, compounds 16-1 to 16-2, having the following formula, were also prepared:

Ex. R⁶ 16-1 —CH₂CH(CH₃)₂ 16-2 benzyl

-   (16-1)     2-{6-[6-((R)-1-mercaptomethyl-3-methylbutylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]-pyridin-3-yl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₃₁H₃₆N₄O₃S, 545.25. found 545.6. -   (16-2)     2-{6-[6-((R)-1-benzyl-2-mercaptoethylcarbamoyl)-4-methyl-2-propyl-benzoimidazol-1-ylmethyl]-pyridin-3-yl}benzoic     acid. MS m/z: [M+H⁺] calcd for C₃₄H₃₄N₄O₃S, 579.24. found 579.4.

ASSAY 1 AT₁ and AT₂ Radioligand Binding Assays

These in vitro assays were used to assess the ability of test compounds to bind to the AT₁ and the AT₂ receptors.

Membrane Preparation From Cells Expressing Human AT₁ or AT₂ Receptors

Chinese hamster ovary (CHO—K1) derived cell lines stably expressing the cloned human AT₁ or AT₂ receptors, respectively, were grown in HAM's-F12 medium supplemented with 10% fetal bovine serum, 10 μg/ml penicillin/streptomycin, and 500 μg/ml geneticin in a 5% CO₂ humidified incubator at 37° C. AT₂ receptor expressing cells were grown in the additional presence of 100 nM PD123,319 (AT₂ antagonist). When cultures reached 80-95% confluence, the cells were washed thoroughly in PBS and lifted with 5 mM EDTA. Cells were pelleted by centrifugation and snap frozen in MeOH-dry ice and stored at −80° C. until further use.

For membrane preparation, cell pellets were resuspended in lysis buffer (25 mM Tris/HCl pH 7.5 at 4° C., 1 mM EDTA, and one tablet of Complete Protease Inhibitor Cocktail Tablets with 2 mM EDTA per 50 mL buffer (Roche cat.#1697498, Roche Molecular Biochemicals, Indianapolis, Ind.)) and homogenized using a tight-fitting Dounce glass homogenizer (10 strokes) on ice. The homogenate was centrifuged at 1000×g, the supernatant was collected and centrifuged at 20,000×g. The final pellet was resuspended in membrane buffer (75 mM Tris/HCl pH 7.5, 12.5 mM MgCl₂, 0.3 mM EDTA, 1 mM EGTA, 250 mM sucrose at 4° C.) and homogenized by extrusion through a 20G gauge needle. Protein concentration of the membrane suspension was determined by the method described in Bradford (1976) Anal Biochem. 72:248-54. Membranes were snap frozen in MeOH-dry ice and stored at −80° C. until further use.

Ligand Binding Assay to Determine Compound Affinities for the Human AT₁ and AT₂ Angiotensin Receptors

Binding assays were performed in 96-well Acrowell filter plates (Pall Inc., cat.#5020) in a total assay volume of 100 μL with 0.2 μg membrane protein for membranes containing the human AT₁ receptor, or 2 μg membrane protein for membranes containing the human AT₂ receptor in assay buffer (50 mM Tris/HCl pH 7.5 at 20° C., 5 mM MgCl₂, 25 μM EDTA, 0.025% BSA). Saturation binding studies for determination of K_(d) values of the ligand were done using N-terminally Europium-labeled angiotensin-II ([Eu]AngII, H-(Eu—N¹)-Ahx-Asp-Arg-Val-Tyr-11e-His-Pro-Phe-OH; PerkinElmer, Boston, Mass.) at 8 different concentrations ranging from 0.1 nM to 30 nM. Displacement assays for determination of pK_(i) values of test compounds were done with [Eu]AngII at 2 nM and 11 different concentrations of drug ranging from 1 μM to 10 μM. Drugs were dissolved to a concentration of 1 mM in DMSO and from there serially diluted into assay buffer. Nonspecific binding was determined in the presence of 10 μM unlabeled angiotensin-II. Assays were incubated for 120 minutes in the dark, at room temperature or 37° C., and binding reactions were terminated by rapid filtration through the Acrowell filter plates followed by three washes with 200 μL ice cold wash buffer (50 mM Tris/HCl pH 7.5 at 4° C., 5 mM MgCl₂) using a Waters filtration manifold. Plates were tapped dry and incubated with 50 μl DELFIA Enhancement Solution (PerkinElmer cat.#4001-0010) at room temperature for 5 minutes on a shaker. Filter-bound [Eu]AngII was quantitated immediately on a Fusion plate reader (PerkinElmer) using Time Resolved Fluorescence (TRF). Binding data were analyzed by nonlinear regression analysis with the GraphPad Prism Software package (GraphPad Software, Inc., San Diego, Calif.) using the 3-parameter model for one-site competition. The BOTTOM (curve minimum) was fixed to the value for nonspecific binding, as determined in the presence of 10 μM angiotensin II. K₁ values for drugs were calculated from observed IC₅₀ values and the K_(d) value of [Eu]AngII according to the Cheng-Prusoff equation described in Cheng et al. (1973) Biochem Pharmacol. 22(23):3099-108. Selectivities of test compounds for the AT₁ receptor over the AT₂ receptor were calculated as the ratio of AT₂K_(i)/AT₁K_(i). Binding affinities of test compounds were expressed as negative decadic logarithms of the Ic values (pK_(i)).

In this assay, a higher pK_(i) value indicates that the test compound has a higher binding affinity for the receptor tested. Exemplary compounds of the invention that were tested in this assay, typically were found to have a pK_(i) at the AT₁ receptor greater than or equal to about 5.0. For example, the compound of Example 1 was found to have a plc value greater than about 7.0.

ASSAY 2 In Vitro Assays for the Quantitation of Inhibitor Potencies (IC₅₀) at Human and Rat NEP, and Human ACE

The inhibitory activities of compounds at human and rat NEP and human ACE were determined using in vitro assays as described below.

Extraction of NEP Activity from Rat Kidneys

Rat NEP was prepared from the kidneys of adult Sprague Dawley rats. Whole kidneys were washed in cold PBS and brought up in ice-cold lysis buffer (1% Triton X-114, 150 mM NaCl, 50 mM Tris pH 7.5; Bordier (1981) J. Biol. Chem. 256:1604-1607) in a ratio of 5 mL of buffer for every gram of kidney. Samples were homogenized using a polytron hand held tissue grinder on ice. Homogenates were centrifuged at 1000×g in a swinging bucket rotor for 5 minutes at 3° C. The pellet was resuspended in 20 mL of ice cold lysis buffer and incubated on ice for 30 minutes. Samples (15-20 mL) were then layered onto 25 mL of ice-cold cushion buffer (6% w/v sucrose, 50 mM pH 7.5 Tris, 150 mM NaCl, 0.06%, Triton X-114), heated to 37° C. for 3-5 minutes and centrifuged at 1000×g in a swinging bucket rotor at room temperature for 3 minutes. The two upper layers were aspirated off, leaving a viscous oily precipitate containing the enriched membrane fraction. Glycerol was added to a concentration of 50% and samples were stored at −20° C. Protein concentrations were quantitated using a BCA detection system with BSA as a standard.

Enzyme Inhibition Assays

Recombinant human NEP and recombinant human ACE were obtained commercially (R&D Systems, Minneapolis, Minn., catalog numbers 1182-ZN and 929-ZN, respectively). The fluorogenic peptide substrate Mca-BK2 (Mca-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-Lys(Dnp)-OH; Johnson et al. (2000) Anal. Biochem. 286: 112-118) was used for the human NEP and ACE assays, and Mca-RRL (Mca-DArg-Arg-Leu-(Dnp)-OH; Medeiros et al. (1997) Braz. J. Med. Biol. Res. 30:1157-1162) was used for the rat NEP assay (both from Anaspec, San Jose, Calif.).

The assays were performed in 384-well white opaque plates at room temperature using the respective fluorogenic peptides at a concentration of 10 μM in assay buffer (50 mM Tris/HCl at 25° C., 100 mM NaCl, 0.01% Tween-20, 1 μM Zn, 0.025% BSA). Human NEP and human ACE were used at concentrations that resulted in quantitative proteolysis of 5 μM of Mca-BK2 within 20 minutes at room temperature. The rat NEP enzyme preparation was used at a concentration that yielded quantitative proteolysis of 3 μM of Mca-RRL within 20 minutes at room temperature.

Assays were started by adding 25 μL, of enzyme to 12.5 μL, of test compound at 12 concentrations (10 μM to 20 μM). Inhibitors were allowed to equilibrate with the enzyme for 10 minutes before 12.5 μL, of the fluorogenic substrates were added to initiate the reaction. Reactions were terminated by the addition of 10 μL, of 3.6% glacial acetic acid after 20 minutes of incubation. Plates were read on a fluorometer with excitation and emission wavelengths set to 320 nm and 405 nm, respectively.

Raw data (relative fluorescence units) were normalized to % activity from the average high readings (no inhibition, 100% enzyme activity) and average low readings (full inhibition, highest inhibitor concentration, 0% enzyme activity) using three standard NEP and ACE inhibitors, respectively. Nonlinear regression of the normalized data was performed using a one site competition model (GraphPad Software, Inc., San Diego, Calif.). Data were reported as plC₅₀ values.

Exemplary compounds of the invention that were tested in this assay, typically were found to have a plC₅₀ for the NEP enzyme greater than or equal to about 5.0, for example, the compound of Example 1 has a plC₅₀ value greater than or equal to about 7.0.

ASSAY 3 Pharmacodynamic (PD) Assay for ACE, AT₁, and NEP Activity in Anesthetized Rats

Male, Sprague Dawley, normotensive rats are anesthetized with 120 mg/kg (i.p.) of inactin. Once anesthetized, the jugular vein, carotid artery (PE 50 tubing) and bladder (URI-1 urinary silicone catheter) are cannulated and a tracheotomy is performed (Teflon Needle, size 14 gauge) to faciliate spontaneous respiration. The animals are then allowed a 60 minute stablization period and kept continuously infused with 5 mL/kg/h of saline (0.9%) throughout, to keep them hydrated and ensure urine production. Body temperature is maintained throughout the experiment by use of a heating pad. At the end of the 60 minute stabilization period, the animals are dosed intravenously (i.v.) with two doses of angiotensin (AngI, 1.0 μg/kg, for ACE inhibitor activity; AngII, 0.1 μg/kg, for AT₁ receptor antagonist activity) at 15 minutes apart. At 15 minutes post-second dose of angiotensin (AngI or AngII), the animals are treated with vehicle or test compound. Five minutes later, the animals are additionally treated with a bolus i.v. injection of atrial natriuretic peptide (ANP; 30 μg/kg). Urine collection (into pre-weighted eppendorf tubes) is started immediately after the ANP treatment and continued for 60 minutes. At 30 and 60 minutes into urine collection, the animals are re-challenged with angiotensin (AngI or AngII). Blood pressure measurements are done using the Notocord system (Kalamazoo, Mich.). Urine samples are frozen at −20° C. until used for the cGMP assay. Urine cGMP concentrations are determined by Enzyme Immuno Assay using a commercial kit (Assay Designs, Ann Arbor, Mich., Cat. No. 901-013). Urine volume is determined gravimetrically. Urinary cGMP output is calculated as the product of urine output and urine cGMP concentration. ACE inhibition or AT₁ antagonism is assessed by quantifying the % inhibition of pressor response to AngI or AngII, respectively. NEP inhibition is assessed by quantifying the potentiation of ANP-induced elevation in urinary cGMP output.

ASSAY 4 In Vivo Evaluation of Antihypertensive Effects in the Conscious SHR Model of Hypertension

Spontaneously hypertensive rats (14-20 weeks of age) are allowed a minimum of 48 hours acclimation upon arrival at the testing site. Seven days prior to testing, the animals are either placed on a restricted low-salt diet with food containing 0.1% of sodium for sodium depleted SHRs (SD-SHR) or are placed on a normal diet for sodium repleted SHRs (SR—SHR). Two days prior to testing, the animals are surgically implemented with catheters into a carotid artery and the jugular vein (PESO polyethylene tubing) connected via a PE10 polyethylene tubing to a selected silicone tubing (size 0.020 ID×0.037 OD×0.008 wall) for blood pressure measurement and test compound delivery, respectively. The animals are allowed to recover with appropriate post operative care.

On the day of the experiment, the animals are placed in their cages and the catheters are connected via a swivel to a calibrated pressure transducer. After 1 hour of acclimation, a baseline measurement is taken over a period of at least 5 minutes. The animals are then dosed i.v. with vehicle or test compound in ascending cumulative doses every 60 minutes followed by a 0.3 mL saline to clear the catheter after each dose. Data is recorded continuously for the duration of the study using Notocord software (Kalamazoo, Mich.) and stored as electronic digital signals. In some studies, the effects of a single i.v. or oral (gavage) dose are monitored for at least 6 hours after dosing. Parameters measured are blood pressure (systolic, diastolic and mean arterial pressure) and heart rate.

ASSAY 5 In Vivo Evaluation of Antihypertensive Effects in the Conscious DOCA-Salt Rat Model of Hypertension

CD rats (male, adult, 200-300 grams, Charles River Laboratory, USA) are allowed a minimum of 48 hours acclimation upon arrival at the testing site before they are placed on a high salt diet.

One week after the start of the high salt diet, a DOCA-salt pellet (100 mg, 21 days release time, Innovative Research of America, Sarasota, Fla.) is implanted subcutaneously and unilateral nephrectomy is performed. On 16 or 17 days post DOCA-salt pellet implantation, animals are implanted surgically with catheters into a carotid artery and the jugular vein with a PESO polyethylene tubing, which in turn was connected via a PE10 polyethylene tubing to a selected silicone tubing (size 0.020 ID×0.037 OD×0.008 wall) for blood pressure measurement and test compound delivery, respectively. The animals are allowed to recover with appropriate post operative care.

On the day of the experiment, each animal is kept in its cage and connected via a swivel to a calibrated pressure transducer. After 1 hour of acclimation, a baseline measurement is taken over a period of at least five minutes. The animals are then dosed i.v. with a vehicle or test compound in escalating cumulative doses every 60 minutes followed by 0.3 mL of saline to flush the catheter after each dose. In some studies, the effects of a single intravenous or oral (gavage) dose is tested and monitored for at least 6 hours after dosing. Data is recorded continuously for the duration of the study using Notocord software (Kalamazoo, Mich.) and stored as electronic digital signals. Parameters measured are blood pressure (systolic, diastolic and mean arterial pressure) and heart rate. For cumulative and single dosing, the percentage change in mean arterial pressure (MAP, mmHg) or heart rate (HR, bpm) is determined as described for Assay 4.

While the present invention has been described with reference to specific aspects or embodiments thereof, it will be understood by those of ordinary skilled in the art that various changes can be made or equivalents can be substituted without departing from the true spirit and scope of the invention. Additionally, to the extent permitted by applicable patent statues and regulations, all publications, patents and patent applications cited herein are hereby incorporated by reference in their entirety to the same extent as if each document had been individually incorporated by reference herein. 

1. A compound of formula I:

wherein: r is 0, 1 or 2; Ar is an aryl group selected from:

R¹ is selected from —COOR″, —NHSO₂R^(1b), —SO₂NHR^(1d), —SO₂OH, —C(O)NH—SO₂R^(1c), —P(O)(OH)₂, —CN, —OCH(R^(1e))—COOH, tetrazol-5-yl,

 where R^(1a) is H, —C₁₋₆alkyl, —C₁₋₃alkylenearyl, —C₁₋₃alkyleneheteroaryl, —C₃₋₇cycloalkyl, —CH(C₁₋₄alkyl)OC(O)R¹″, —C₀₋₆alkylenemorpholine,

 R^(1aa) is —O—C₁₋₆alkyl, —O—C₃₋₇cycloalkyl, —NR^(1ab)R^(1ac), or —CH(NH₂)CH₂COOCH₃; R^(1ab) and R^(1ca) are independently selected from H, —C₁₋₆alkyl, and benzyl, or are taken together as —(CH₂)₃₋₆—; R^(1b) is R^(1c) or —NHC(O)R^(1c); R^(1c) is —C₁₋₆alkyl, —C₀₋₆alkylene-O—R^(1ca), —C₁₋₅alkylene-NR^(1cb)R^(1cc), —C₀₋₄alkylenearyl or —C₀₋₄alkyleneheteroaryl; R^(1ca) is H, —C₁₋₆alkyl, or —C₁₋₆alkylene-O—C₁₋₆alkyl; R^(1cb) and R^(1cc) are independently selected from H and —C₁₋₆alkyl, or are taken together as —(CH₂)₂—O—(CH₂)₂— or —(CH₂)₂—N[C(O)CH₃]-(CH₂)₂—; R^(1d) is H, R^(1c), —C(O)R^(1c), or —C(O)NHR^(1c); R^(1c) is —C₁₋₄alkyl or aryl; n is 0, 1, 2 or 3; each R² is independently selected from halo, —NO₂, —C₁₋₆alkyl, —C₂₋₆alkenyl, —C₃₋₆cycloalkyl, —CN, —C(O)R^(2a), —C₀₋₅alkylene-OR^(2b), —C₀₋₅alkylene-NR^(2c)R^(2d), —C₀₋₃alkylenearyl, and —C₀₋₃alkyleneheteroaryl; where R^(ea) is H, —C₁₋₆alkyl, —C₃₋₆cycloalkyl, —OR^(2b), or —NR^(2c)R^(2d); R^(2b) is H, —C₁₋₆alkyl, —C₃₋₆cycloalkyl, or —C₀₋₁alkylenearyl; and R^(2c) and R^(ed) are independently selected from H, —C₁₋₄alkyl, and —C₀₋₁alkylenearyl; R³ is selected from —C₁₋₁₀alkyl, —C₂₋₁₀alkenyl, —C₃₋₁₀alkynyl, —C₀₋₃alkylene-C₃₋₇cycloalkyl, —C₂₋₃alkenylene-C₃₋₇cycloalkyl, —C₂₋₃alkynylene-C₃₋₇cycloalkyl, —C₀₋₅alkylene-NR^(3a)—C₀₋₅alkylene-R^(3b), —C₀₋₅alkylene-O—C₀₋₅alkylene-R^(3b), —C₀₋₅alkylene-S—C₁₋₅alkylene-R^(3b), and —C₀₋₃alkylenearyl; where R^(1a) is H, —C₁₋₆alkyl, —C₃₋₇cycloalkyl, or —C₀₋₃alkylenearyl; and R^(3b) is H, —C₁₋₆alkyl, —C₃₋₇cycloalkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl, or aryl; X is —C₁₋₁₂alkylene-, where at least one —CH₂— moiety in the alkylene is replaced with a —NR^(4a)—C(O)— or —C(O)—NR^(4a)— moiety, where R^(4a) is H, —OH, or —C₁₋₄alkyl; R⁵ is selected from —C₀₋₃alkylene-SR^(5a), —C₀₋₃alkylene—C(O)NR^(5b)R^(5c), —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d), —NH—C₀₋₁alkylene-P(O)(OR^(5e))₂, —C₀₋₃alkylene-P(O)OR^(5e)R^(5f), —C₀₋₂alkylene-CHR^(5g)—COOH, —C₀₋₃alkylene—C(O)NR^(5l)—CHR^(5l)—COOH, and —C₀₋₃alkylene-S—SR^(5j); where R^(5a) is H or —C(O)—R^(5aa); R^(5aa) is —C₁₋₆alkyl, —C₀₋₆alkylene-C₃₋₇cycloalkyl, -aminoC₄₋₇cycloalkyl, —C₀₋₆alkylenearyl, —C₀₋₆alkyleneheteroaryl, —C₀₋₆alkylenemorpholine, —C₀₋₆alkylenepiperazine-CH₃, —C₀₋₆alkylenepiperidine, —C₀₋₆alkylenepiperidine—CH₃, —CH[N(R^(5ab))₂]-aa where aa is an amino acid side chain, —C₀₋₆alkylene-CH[N(R^(5ab))₂]—R^(5aa), -2-pyrrolidine, -2-tetrahydrofuran, —C₀₋₆alkylene-OR^(5ab), —O—C₀₋₆alkylenearyl, —C₁₋₂alkylene-OC(O)—C₁₋₆alkyl, —C₁₋₂alkylene-OC(O)—C₀₋₆alkylenearyl, —O—C₁₋₂alkylene-OC(O)O—C₁₋₆alkyl, —C₋₄alkylene-COOH, or -arylene-COOH; R^(5ab) is independently H or —C₁₋₆alkyl; R^(5ac) is H, —C₁₋₆alkyl, —CH₂—C₃₋₇cycloalkyl or —COOH; R^(5b) is H, —OH, —OC(O)R^(5ba), —CH₂COOH, —O-benzyl, -pyridyl, or —OC(S)NR^(5bb)R^(5bc); R^(5ba) is H, —C₁₋₆alkyl, aryl, —OCH₂-aryl, —CH₂O-aryl, or —NR^(5bb)R^(5bc); R^(5bb) and R^(5b)′ are independently selected from H and —C₁₋₄alkyl; R^(5c) is H, —C₁₋₆alkyl, or —C(O)R⁵″; R^(5aa) is —C₁₋₆alkyl, —C₃₋₇cycloalkyl, aryl, or heteroaryl; R^(5d) is H, —C₁₋₄alkyl, —C₀₋₃alkylenearyl, —NR^(5da)R^(5db), —CH₂SH, or —O—C₁₋₆alkyl; R^(5da) and R^(5db) are independently selected from H and —C₁₋₄alkyl; R^(5e) is H, —C₁₋₆alkyl, —C₁₋₃alkylenearyl, —C₁₋₃alkyleneheteroaryl, —C₃₋₇cycloalkyl, —CH(CH₃)OC(O)R^(5ea),

 R^(5aa) is —O—C₁₋₆alkyl, —O—C₃₋₇cycloalkyl, —NR^(5eb)R^(5ec), or —CH(NH₂)CH₂COOCH₃; R^(5eb) and R^(5aa) are independently selected from H, —C₁₋₄alkyl, and —C₁₋₃alkylenearyl, or are taken together as —(CH₂)₃₋₆—; R^(5f) is H, —C₁₋₄alkyl, —C₀₋₃alkylenearyl, —C₁₋₃alkylene-NR^(5fa)R^(5fb), or —C₁₋₃alkylene(aryl)-C₀₋₃alkylene-NR^(5fa)R^(5fb); R^(5fa) and R^(5fb) are independently selected from H and —C₁₋₄alkyl; R^(5g) is H, —C₁₋₆alkyl, —C₁₋₃alkylenearyl, or —CH₂—O—(CH₂)₂—OCH₃; R^(5h) is H or —C₁₋₄alkyl; R^(5i) is H, —C₁₋₄alkyl, or —C₀₋₃alkylenearyl; and R^(5j) is —C₁₋₆alkyl, aryl, or —CH₂CH(NH₂)COOH; R⁶ is selected from —C₁₋₆alkyl, —CH₂—O—(CH₂)₂OCH₃, —C₁₋₆alkylene-O—C₁₋₆alkyl, —C₀₋₃alkylenearyl, —C₀₋₃alkyleneheteroaryl, and —C₀₋₃alkylene-C₃₋₇cycloalkyl; and R⁷ is H or is taken together with R⁶ to form —C₃₋₈cycloalkyl; wherein: each —CH₂— group in —(CH₂)_(r)— is optionally substituted with 1 or 2 substituents independently selected from —C₁₋₄alkyl and fluoro; each carbon atom in the alkylene moiety in X is optionally substituted with one or more R^(4b) groups and one —CH₂— moiety in X may be replaced with a group selected from —C₃₋₈cycloalkylene-, —CR^(4d)═CH—, and —CH═CR^(4d)—; where R^(4b) is —C₀₋₅alkylene-COOR^(4c), —C₁₋₆ alkyl, —C₀₋₁alkylene-CONH₂, —C₁₋₂ alkylene-OH, —C₀₋₃alkylene-C₃ ₋₇ cycloalkyl, 1H-indol-3-yl, benzyl, or hydroxybenzyl; R^(4c) is H or —C₁₋₄alkyl; and R^(od) is —CH₂-thiophene or phenyl; each alkyl and each aryl in R¹⁻³, R^(4a-4d), and R⁵⁻⁶ is optionally substituted with 1 to 7 fluoro atoms; Ar and each aryl and heteroaryl in R¹⁻³ and R⁵⁻⁶ is optionally substituted with 1 to 3 substituents independently selected from —OH, —C₁₋₆alkyl, —C₂₋₄alkenyl, —C₂₋₄alkynyl, —CN, halo, —O—C₁₋₆ alkyl, —S—C₁₋₆ alkyl, —S(O)—C₁₋₆ alkyl, —S(O)₂—C₁₋₄alkyl, -phenyl, —NO₂, —NH₂, —NH—C₁₋₆alkyl and —N(C₁₋₆alkyl)₂, wherein each alkyl, alkenyl and alkynyl is optionally substituted with 1 to 5 fluoro atoms; and pharmaceutically acceptable salts thereof.
 2. The compound of claim 1, wherein r is
 1. 3. The compound of claim 1, wherein Ar

and Ar is optionally substituted with one fluoro, chloro or bromo atom.
 4. The compound of claim 1, wherein R¹ is selected from —COOH, —NHSO₂R^(1b), —SO₂NHR^(1d), —SO₂OH, —C(O)NH—SO₂R^(1c), —P(O)(OH)₂, —CN, —O—CH(R^(1e))—COOH, tetrazol-5-yl,


5. The compound of claim 4, wherein R¹ is selected from —COOH, —SO₂NHR^(1d), and tetrazol-5-yl.
 6. The compound of claim 1, wherein R¹ is —COOR^(1a) and R^(1a) is selected from —C₁ ₋₆ alkyl, —C₁ ₋₃ alkylenearyl, —C₁ ₋₃ alkylenehetero aryl, —C₃₋₇ cyclo alkyl, —CH(C₁₋₄alkyl)OC(O)R¹″, —C₀₋₆alkylenemorpholine,


7. The compound of claim 1, wherein n is 0 or
 1. 8. The compound of claim 7, wherein n is 1 and R² is at the 4 position of the benzoimidazole ring.
 9. The compound of claim 1, wherein R² is selected from halo, —C₁₋₆alkyl, and —C₀₋₅alkylene-OR^(2b).
 10. The compound of claim 9, wherein R² is selected from chloro, —CH₃, —CH₂F, —CF₃, and —CH₂OH.
 11. The compound of claim 1, wherein R³ is selected from —C₁₋₁₀alkyl and —C₀₋₅alkylene-O—C₀₋₅alkylene-R^(3b), and R^(ab) is —C₁₋₆alkyl.
 12. The compound of claim 11, wherein R³ is —C₂₋₅alkyl.
 13. The compound of claim 1, wherein X is —C₁₋₆alkylene- and one to four —CH₂— moieties are replaced with a —NR^(4a)—C(O)— or —C(O)—NR^(4a)— moiety.
 14. The compound of claim 13, wherein X is selected from —C(O)NH—, —NHC(O)—, and —CH₂—NHC(O)—.
 15. The compound of claim 1, wherein R⁵ is selected from —C₀₋₃alkylene-SR^(5a), —C₀₋₃alkylene—C(O)NR^(5b)R^(5c), —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d), —NH—C₀₋₁alkylene-P(O)(OR^(5e))₂, —C₀₋₃alkylene-P(O)OR^(5e)R^(5f), —C₀₋₂alkylene-CHR^(5g)—COOH, and —C₀₋₃alkylene—C(O)NR^(5h)—CHR^(5l)—COOH; R^(5a) is H; R^(5b) is —OH; R^(5a) is H; R^(5d) is H; and R^(5e) is H.
 16. The compound of claim 1, wherein R⁵ is selected from —C₀₋₃alkylene-SR^(5a), —C₀₋₃alkylene—C(O)NR^(5b)R^(5c), —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d), —NH—C₀₋₁alkylene-P(O)(OR^(5e))₂, —C₀₋₃alkylene-P(O)OR^(5e)R^(5f), and —C₀₋₃alkylene-S—SR^(5j); R^(5a) is —C(O)—R^(5aa); R^(5b) is H, —OC(O)R^(5ba), —CH₂COOH, —O-benzyl, -pyridyl, or —OC(S)NR^(5bb)R^(5bc); and R^(5e) is —C₁₋₆alkyl, —C₁₋₃alkylenearyl, —C₁₋₃alkylenehetero aryl, —C₃₋₇cyclo alkyl, —CH(CH₃)—O—C(O)R^(5ea),


17. The compound of claim 15, wherein R¹ is selected from —COOH, —NHSO₂R^(1b), —SO₂NHR^(1d), —SO₂OH, —C(O)NH—SO₂R^(1c), —P(O)(OH)₂, —CN, —O—CH(R^(1e))—COOH, tetrazol-5-yl,


18. The compound of claim 16, wherein R¹ is —COOR^(1a), and R^(1a) is selected from —C₁₋₆alkyl, —C₁₋₃alkylenearyl, —C₁₋₃alkylenehetero aryl, —C₃₋₇cyclo alkyl, —CH(C₁₋₄alkyl)-OC(O)R^(1aa), —C₀₋₆alkylenemorpholine,


19. The compound of claim 15, wherein R¹ is —COOR^(1a), and R^(1a) is selected from —C₁₋₆alkyl, —C₁₋₃alkylenearyl, —C₁₋₃alkylenehetero aryl, —C₃₋₇cycloalkyl, —CH(C₁₋₄alkyl)-OC(O)R^(1aa), —C₀₋₆alkylenemorpholine,


20. The compound of claim 16, wherein R¹ is selected from —COOH, —NHSO₂R^(1b), —SO₂NHR^(1d), —SO₂OH, —C(O)NH—SO₂R^(1c), —P(O)(OH)₂, —CN, —O—CH(R^(1e))—COOH, tetrazol-5-yl,


21. The compound of claim 1, wherein R⁶ is selected from —C₁₋₆alkyl, —C₀₋₃alkylenearyl, and —C₀₋₃alkylene-C₃₋₇cycloalkyl.
 22. The compound of claim 1, wherein R⁷ is H. 23-29. (canceled)
 30. The compound of claim 1, having formula Ib:


31. The compound of claim 30, wherein: r is 1; R¹ is —COOR^(1a), and R^(1a) is H or —C₁₋₆alkyl; n is 1; R² is —C₁₋₆alkyl; R³ is —C₁₋₁₀alkyl; X is selected from —CH₂—NHC(O)—, —NHC(O)—, and —C(O)NH—; R⁵ is selected from —C₀₋₃alkylene-SR^(5a), —C₀₋₃alkylene—C(O)—NR^(5b)R^(5c), and —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d); R^(5a) is H or —C(O)—R^(5aa); R^(5aa) is —C₁₋₆alkyl; R^(5b) is —OH; R⁵ is H; R^(5d) is H or —C₁₋₄alkyl; R⁶ is selected from —C₁₋₆alkyl, —C₀₋₃alkylenearyl, and —C₀₋₃alkylene-C₃₋₇cycloalkyl; R⁷ is H; the alkyl in R⁶ is optionally substituted with 1 to 7 fluoro atoms; and the phenyl ring is optionally substituted with 1 halo atom.
 32. The compound of claim 30, wherein X is —CH₂—NHC(O)—.
 33. The compound of claim 32, wherein: r is 1; R¹ is —COOR^(1a), and R^(1a) is H or —C₁₋₆alkyl; n is 1; R² is —C₁₋₆alkyl; R³ is —C₁₋₁₀alkyl; R⁵ is selected from —C₀₋₃alkylene-SR^(5a) and —C₀₋₃alkylene—C(O)NR^(5b)R^(5c); R^(5a) is H or —C(O)—R^(5aa); R^(5aa) is —C₁₋₆alkyl; R^(5b) is —OH; R⁵ is H; R⁶ is selected from —C₁₋₆alkyl, —C₀₋₃alkylenearyl, and —C₀₋₃alkylene-C₃₋₇cycloalkyl; R⁷ is H; the alkyl in R⁶ is optionally substituted with 1 to 7 fluoro atoms; and the phenyl ring is optionally substituted with 1 halo atom.
 34. The compound of claim 30, wherein X is —C(O)NH—.
 35. The compound of claim 34, wherein: r is 1; R¹ is —COOR^(1a), where R^(1a) is H or —C₁₋₆alkyl; n is 1; R² is —C₁₋₆alkyl; R³ is —C₁₋₁₀alkyl; R⁵ is selected from —C₀₋₃alkylene-SR^(5a), —C₀₋₃alkylene—C(O)NR^(5b)R^(5c), and —C₀₋₃alkylene-NR^(5b)—C(O)R^(5d); R^(5a) is H or —C(O)—R^(5aa); R^(5aa) is —C₁₋₆alkyl; R^(5b) is —OH; R⁵ is H; R^(5d) is H or —C₁₋₄alkyl; R⁶ is selected from —C₁₋₆alkyl and —C₀₋₃alkylenearyl; and R⁷ is H. 36-39. (canceled)
 40. A pharmaceutical composition comprising a compound of claim 1 and a pharmaceutically acceptable carrier.
 41. The pharmaceutical composition of claim 40, further comprising a second therapeutic agent selected from diuretics, β₁ adrenergic receptor blockers, calcium channel blockers, angiotensin-converting enzyme inhibitors, AT₁ receptor antagonists, neprilysin inhibitors, non-steroidal anti-inflammatory agents, prostaglandins, anti-lipid agents, anti-diabetic agents, anti-thrombotic agents, renin inhibitors, endothelin receptor antagonists, endothelin converting enzyme inhibitors, aldosterone antagonists, angiotensin-converting enzyme/neprilysin inhibitors, vasopressin receptor antagonists, and combinations thereof.
 42. (canceled)
 43. The compound of claim 1 prepared by the process comprising: (a) coupling a compound of formula 1 with a compound of formula 2:

 to produce a compound having the formula:

 where: A is —NH₂ and B is —COOH, or A is —COOH and B is —NH₂; the sum of a and b is in the range of 0 to 11; Ar* is Ar—R¹*, where R¹* is R¹ or a protected form of R¹; and R⁵* is R⁵ or a protected form of R⁵; the carbon atoms in the —(CH_(2a) and —(CH₂)_(b) groups may be substituted with one or more R^(4b) groups; and one —CH₂— group in the —(CH_(2a) or the —(CH_(2b) group may be replaced with —C₃₋₈cycloalkylene-, —CR^(4d)═CH—, or —CH═CR^(4d)—; and (b) when R¹* is a protected form of R¹ and/or R⁵* is a protected form of R⁵, deprotecting the product of step (a) to produce a compound of formula I.
 44. An intermediate useful in the synthesis of a compound of claim 1, selected from:

where Ar* is Ar—R¹*; R¹* is selected from —C(O)O—P², —SO₂O—P⁵, —SO₂NH—P⁶, —P(O)(O—P⁷)₂, —OCH(R^(1e))—C(O)O—P², and tetrazol-5-yl-P⁴; R⁵* is selected from —C₀₋₃alkylene-S—P³, —C₀₋₃alkylene—C(O)NH(O—P⁵), —C₀₋₃alkylene-N(O—P⁵)—C(O)R^(5d), —C₀₋₁alkylene-NHC(O)CH₂S—P³, —NH—C₀₋₁alkylene-P(O)(O—P⁷)₂, —C₀₋₃alkylene-P(O)(O—P⁷)—R^(5f), —C₀₋₂alkylene-CHR^(5g)—C(O)O—P², and —C₀₋₃alkylene—C(O)NR^(5h)—CHR^(5l)—C(O)O—P²; P² is a carboxy-protecting group; P³ is a thiol-protecting group; P⁴ is a tetrazole-protecting group; P⁵ is a hydroxyl-protecting group; P⁶ is a sulfonamide-protecting group; and P⁷ is a phosphonate-protecting group or phosphinate-protecting group; and salts thereof.
 45. The intermediate of claim 44, selected from the group comprising:

46-50. (canceled) 